524681 經濟部中央標準局員工消費合作社印製 Α7 Β7 五、發明説明（1 ) 發明背景 本申請案傜為各於1993.3· 16·和1990 ·8.13.提申的共 同在審查中的專利申請案（copending applicaton)序列编 號0.8/033, 105和07/566,996之一部分延續申請案（contin-uation-in-part)，而後者復為19δ9·δ·14·提申而現已放棄 的申請案序列编號07/394,377之一部分延續申請案。 本發明一般而言傜有闊於外科儀器與外科技術。更明 * 確地說，本發明偽針對一用以經由一位於眼内之標準尺寸 大的切口來移植呈一渦形構形之視網膜细胞之薄片、上皮 组織和/或脈絡膜組織的外科工具，一供移植至眼之視網 膜下區域之移植物以及一用以重建營養不良的視網膜、視 網膜色素上皮層及脈絡膜之方法。 視網膜傜為感覺上皮表面，其襯於眼後房，接收由晶 體所形成之影像，將此影像轉換成神經脈衝並藉由視神經 將此訊息傳輸至腦。視網膜包含有數層，亦即神經節細胞 層、内網織層、内穎粒層、外網織層、外顆粒層、光感受 器内節段與外節段。該外穎粒層包含有感光細胞之細胞體 ，其帶有該内及外節段成為該細胞體之延伸物。 脈絡膜偽為一位在脊椎動物眼之視網膜與鞏膜外被之 間並含有大量分枝的色素細胞之血管膜。直接位在脈絡膜 與視網膜之間的是視網膜色素上皮，其與感光細胞形成一 密切的結構上與功能上關偽。 數種型式之失明主要偽關係到由視網膜、視網膜色素 上皮、脈络膜内之缺陷或可能的其他因子（例如強光、視 一 5 — 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ297公釐） —--ί------裝-- (請先閲讀背面之注意事項再填寫本頁) 訂 經濟部中央標準局員工消費合作社印製 524681 A7 B7 五、發明説明（2 ) 網膜剝離、眼内出血）所引起的光受體細胞之喪失。在數 種視網膜退化性疾病中，選定的細胞族群會喪失。特別地 ，在斑退化和色素性視網膜炎中，其視網膜光受體會退化 而位於視網膜内之其他細胞以及視網膜的中央接合則被維 持著。在一為恢復早先被認為是一無法修復的受損視網膜 之努力中，研究者曾建議各種不同型式之移揎物與移植技 術，而其無一者能構成一有效型式來重建一營養不良的視 網膜。 將視網膜細胞移植至眼可追溯至Royo et al。，Growt k21:313-336 (1959)之報告，其中胚視網膜被移植至母體 眼之前房。各種不同的細胞，包括光感受霞，被報導會存 活。隨後del Cerro能夠重複並擴展這些實驗（del Cerro e t a 1., Invest. Ophthalmol . Vis. Sci.· 26 «1182-1185, 1985)。緊接而後 Turner et al., Dev . Brain Res , . 26 : 91-104 (1986)顯示新生兒的視網膜組織可被移植至視網 膜傷處内。 在相關的研究中，Simmons et al., Soc . Meurosc i · Abstr, Ιϋ：668 (1984)證實胚視網膜可顱内被移植，存活 下來，顯示出相當正常的發育，能夠神經支配中央結構， 以及以一光-依賴性方式活化這些結構。更甚者，這些顱 内移植物能激發經由宿主的神經条統所調節的光-依賴性 行為反應（瞳孑L反射）0Klassen et a 1 . , Exp. Neurol， 102： 102-108 (1988)以及 Klassen et a 1 · , Proc . Na11 > AnMU. USA 84:6958-6960 (1987)。 一 6 - 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） — 裝 訂 線 (請先閲讀背面之注意事項再填寫本頁) 524681 經濟部中央標準局員工消費合作社印製 A7 B7 五、發明説明（3 ) L ί and Turner , Exp . Eye Res，，47 ♦ 911 (198&)曽 假設利用將視網膜色素上皮（RPE)移植至下視網膜間隙， 而在RCS營養不良的大白鼠體内以其健康的野生型相關對 照物來取代有缺陷的突變型RPE細胞以作為一治療方式。 根據他們的方法，RPE從6至8天大的黑眼大白鼠被分離出 並藉由使用一穿透鞏膜與脈絡膜之損害範例被移植至視網 膜下間隙内。於切口位置處使用一接有一為30號針頭之10 ml注射針將一為lnil的RPE注射物（40,000-60,000細胞）置 入視網膜下間隙。然而，此方法破壞了該移植物所需要的 宿主視網膜色素上皮之細胞極性與天生的组織结構。 del Cerro (del Cerro et a 1 . , Invest. Ophthalmol . Vi^ W.· 21_··11δ2-1185, 1985)報導一用以將一組織條 片移植至前房或宿主視網膜内之方法。該等燦片傺藉由從 供給者眼切下神經視網膜製備而得者。接而將該視網膜切 成適當之組織條片，其隨而利用一 30號針頭或徽吸量管而 被注射到適當的位置内，該條片之寬度被限制至該針頭之 内直徑（25〇μπ)而長度則少於lnun。雖然del Cerro報導視 網膜條片之眼内移植可存活，其注意到該方法有某些一定 的限制。舉例而言，他的技術無法容許就是正在失去的細 胞（例如光受體）之取代，而永遠要包含一由視網膜細胞構 成之混合物。因此，使用這樣一個移植物，不可能做到其 缺乏一持定的細胞族群（例如光受體）之營養不良的視網膜 之適當的再建。 del Cerro e t a 1 ., Neurosc i. Lett。92 »21-26, 一 7 — 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） I 裝 訂 線 (請先閲讀背面之注意事項再填寫本頁) 524681 經濟部中央標準局員工消費合作社印製 A7 B7 五、發明説明（4 ) 1988復報導一用來移植經解離的神經視網膜細胞之方法。 在此方法中，供給者視網膜被切成小片，於胰蛋白酶内培 育15分鐘，並藉由令其通過一精細拉成的吸量管進行抽吸 而將之研製成一單一的細胞懸浮液。相對於上面所討論的 Li and Turner方法，此方法破壞了該移植物（包括供給者 外穎粒層）之有組織化的天生結構。光受體之嚴格的組纖 结構（具有朝向色素上皮之外節段以及面向外網織層之突 篛末端）被喪失了。更甚者，沒有一個用以従其他的視網 膜细胞中分離及純化出任一給定的視網膜細胞族群（例如 光受體）之方法被證明之。 本案發明人相信要恢復一合理程度之視力，將光受體 維持在一有組織的外顆粒層結構内是必要的。此结論傺根 據已詳知的光感受體之光學待徵（外節段發揮有如光導引 ，之作用）以及臨床證據（其顯示視網膜幾何内皺襞或類似的 (甚而徹小的）破裂可嚴重地降解視力。 發明槪要説明 因此，在本發明目的之中，可注意到有一是提供一會 保存一相當大量的從一供給者眼收獲之組織的方法；提供 這樣一種方法，其中一相當大量的經收獲之組織被如此形 ;成而使該經收獲之組織能被插入一位於眼内之標準尺寸大 的切口；提供這樣一種方法，其中該等細胞之極性與組織 結構在收獲之時被維持於移植物内；以及提供一用以將移 植物移植至一眼之視網膜下區域的方法。 進一步地，在本發明的數個目的與持徵之中，可注意 一 8 一 本紙張尺度適用中國國家標準（CNS ) A4規格（21〇X：297公釐） l·--r------裝-- (請先閲讀背面之注意事項再填寫本頁) 、π 線_ 524681 A7 B7 五、發明説明（5 ) 到有一是提供一移植物以供用於營養不良的視網膜之重建 或拯救一罹患一遣傳性或後天性人類視網膜疾病之個體的 内生光受體細胞，該疾病其造成視捍之逐漸喪失而依序地 最终造成視錐之營養不良、功能障礙和/或喪失；.提供這 樣一種移植物，其藉由在移植之時維持光受體之極性結搆 及其與臨近的外網織層之後聯會（postsynaptic)標的的密 切接近而促進光受體軸突之再生長。 次又，在本發明的數値目的與特徵之中，可注意到有 一是提供一供用於該植入方法之外科工具，其形成供插入 一標準尺寸大的切口内之該移植物；以及提供一供用於移 植之外科工具，其容許適當的視網膜局部之置放以及保護 光受體、視纲膜色素上皮組織、脈絡膜組織和/或布魯赫 氏膜（Bruch’s membrane)在該外科裝置被置入眼内之前與 放入之時免受損害。 經濟部中央標準局員工消費合作社印製 ----ί------— 裝-- (請先閱讀背面之注意事項再填寫本頁) 線 一般而言，該植入方法包括盤捲一可植入的物質，該 物質傜為一薄片般形狀而能形成一渦形物者。渦形物之盤 曲彼此不影響另一者俾使該可植入的物質之隨後的解盤得 以大體上回復其原先的薄片般形狀。在宿主眼内做一個切 口以供將渦形物插入一位於宿主眼的視網膜與下層組織之 1間的位置處。該切口較諸將呈解盤的薄片般形狀之該可植 入的物質插入所需之切口要小。首先將該渦形物之一端經 由該切口插入宿主眼至一位於視網膜與下層組織之間的位 置處。在該渦形物之插入後其解盤以呈薄片般之形狀平躺。 於宿主眼的視網膜與下層組織之間且該切口被閉合。 本紙張尺度適用中國國家標準（CNS ) Α4規格（21〇X297公釐） 經濟部中央標準局員工消費合作社印製 524681 A7 __B7 五、發明説明（6 ) 一般而言，該供移植之移植物包含有一層大體上會在 體溫下溶解之無毒的可撓性組成以及一被盤捲以形成一渦 形物之欲予以移植的物質。該渦形物之一端可首先經由切 口（其大小傜依據該渦形物之橫截面區域而定）插入至一供 移植之位置，並接而被解盤以呈薄片般之形狀平躺在該移 植之位置處ϋ . 一般而言，該用以形成一渦形物之植入物包含有一端 開放的並具有一漏斗之管狀體。一載體首先從該管狀體其 開放的一端進入，並經由該漏斗沿其護部來進料。當該載 饅經由該漏斗被進料而與該漏斗的内表面産生之接合導致 該載體盤捲成渦形物。該渦形物之存在始至該漏斗之一狹 窄端。 本發明之其他目的有部分已明顯可知，而有部分將於 下文中予以指明。 圖式槪要說明 第1圖是一如實施例1中所述的正常大白鼠視網膜之一 涪凍切片的相片； 第2圖.是一如實施例1中所述的一經持續的照明後變成 失明的大白鼠視網膜之一冷凍切片的相片； 第3圖是一供給者視網膜之一示意圖； 第4圖是一經整平的視網膜一之示意圖； 第5圖是一被置放在一基質上之經整平的視網膜之一 示意圖； 第6圖是一被置放在一基質上之經切片的視網膜之一 一 10 - 本紙張尺度適用中國國家標準（CNS ) Α4規格（210 X 297公釐） I---r------辦衣------、玎------^ (請先閲讀背面之注意事項再填寫本頁) 經濟部中央標準局員工消費合作社印製 524681 Μ Β7__ 五、發明説明（7 ) 示意圖； 第7圖是一包含有一位在一支撑、安定用的基質上之 視網膜切片的層狀物之一示意圖； 第8圖是第7圖的層狀物之頂端平面示意圖，其顯示一 包含有一光受體細胞層與一支撑、安定用的基質之移植物 (虛線）； 第9圖是被安置在一由間隔物所形成的平板上的層狀 物之一示意圖； 第10圖是被安置在一被注入以明膠加有蓋玻片的平板 上的層狀物之一示意圖； . 第11圖是頂板正被倒向地滑移開之一示意圖； .第12圖是所形成的移植物之一示意圖； 第13圖是正被削切的移植物之一示意圖； 第14圖是該經削切的移植物正從該平板移出以供移植 之一不意圖； 第15圖是一渦形物之一透視圖； 第16圖是一儀器之正視圖，該儀器係供該移植物之盤 捲與植入：而經盤捲之移植物位於該儀器之漏斗内； 第17圖是該儀器之一側視圖，並有一管腔被貼合至儀 器之外側； 第18圖是該儀器之一側視圖，並有一推進該渦形物之 柱塞； 第19圖是一通過一眼之水平剖面圖，其例示一使用該 儀器來部分地延伸橫跨該眼之外科方法； - 11 - ^氏張尺度適用中國國家標準（CNS ) Α4規格（210 X 297公釐） 扣衣 訂 線 (請先閲讀背面之注意事項再填寫本頁) 524681 A 7 B7 五、發明説明（Ο 第20圖是一通過一眼之水平剖面圖，其例示一使用該 儀器插入一水泡内的睫狀體垣部外科方法；. 第21圖是一沿第16圖的線21-22所取的水平剖面圖， 其例示一位於該儀器内之斜坡。 在圖式之數値圖中相關的參考記號意指相關的部分。 薛明詳細説明 如此處所用者，''供給者〃一辭偽指相對於宿主的相 同或相異生物體，而 ''供給者組織〃 一辭係指從獲取自相 對於宿主的相同或相異生物體穫取之組織收獲而得或細胞 培養而得的組織。 經濟部中央標準局員工消費合作社印製 (請先閲讀背面之注意事項再填寫本頁) 數種型式之失明（例如色素性視網膜炎、視網膜脱離 、斑退化和與光暴露有關的失明）主要傜闊係到眼内光受 護之喪失。但是，光受體之破壞並不一定導致剩餘的視網 膜或者聯接視網膜與腦的軸突之喪失。令人驚訝地，本案 發明人發現到藉由以收獲自一供給者且被維持在其原有之 组織結構與細胞極性的光受體來取代受損的光受體，則某 種程度之視力是可被恢復的。更甚者，如進一步載述於共 同在審査中的專利申請案序列编號08/033, 105 (其在此被 併入本案以為參考文獻）中者，收獲自一供給者眼之光受 體視桿之移植可以''拯救〃位於視網膜之内的内生視錐光 受體而因此可恢復或保存仍存在的視錐光受體之視覺敏度 。亦即，本案發明人已發現到經移植的視桿對内生視錐發 揮一營養作用。 第1圖是一正常的大白鼠視網膜之一冷凍切片的相片 一 12 — 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X 297公釐） 經濟部中央標準局員工消費合作社印製 524681 A7 B7___ 五、發明説明（9 ) 。第2圖是一經持續的照明[破隳光受體（外穎粒）層但留下 其他大部分係完整的視網膜層與細胞]的大白鼠視網膜之 一冷凍切片的相片。在這些以及隨後的圖式中，該視網膜 或其層，例如神經節細胞層（"G”）、内網識層（"IPL”）、内 顆粒層（”INL”）、外網織層（”〇PL”）、外穎粒層（”ONL”）、 内節段（"IS”）、外節段（” 〇s”）以及視網膜色素上皮（”R PE”） 從上至下被分別地示出。 現參照第3圖，依據本發明之一方法來製備一供經由 一切口植入的光受體移植物，該切口之寛度較呈薄片般形 狀的該移植物者要小。但是，該移植物可包含有其他可植 入的物質，例如其他的視網膜細胞、抗病毒與抗生物劑和 /或其他的藥理示劑。 藉由從一供給者眼移出一供給者視纲膜50 (包含有内 視纲膜層52與光受體層54)來製備一包含有光受體細胞之 * 移植物。藉由從接近視網膜的中心之位置處至其外緣作數 次的切割（參照第8圔）而將該供給者視網膜50予以整平（第 4圖）。若有需要，亦可在其他方向進行切割。 如第5圖所示的，該經整平的視網膜56以其光受體側54 朝下被置放在一凝膠板58上，該凝膠板之表面已被弄平俾 以提供一與一振動切片機之刀片相平行的平坦表面60。該 明膠板58被固定在該振動切片機之一傳統的夾頭上。將融 ^ 化的4-5%明膠溶液沉積在臨接該經整平的視網膜/明膠表 面之界面61處，並在該經整平的視網膜下方以毛細作用吸 引明膠，致使該經整平的視網膜漂浮在該明膠板58上。迅 一 13 — 本紙張又度適用中國國家標準（CNS ) A4規格（21〇X297公釐） I 裝 訂 線 (請先閲讀背面之注意事項再填寫本頁) 524681 A7 B7 經 濟 部 冲 央 樣 準 局 員 費 合 作 社 印 製 五、發明説明（10) 速地將過量的融化的明膠予以移除，並隨之以冰冷的林格 氏液（R inerts sol lit ion)將該飄浮的、經整平的視網膜冷 卻至約4°C，該林格氏液璟繞明膠塊以造成融化的明膠膠 凝。該經整平的視網膜56因此被貼合至該明膠塊上。 如第6圖所示，該内視網膜部分52從頂端往下以約20 至50ιημ予以切片直到接近光受體層54，於是從該視網膜之 内層（亦即神經節細胞層、内網織層、内穎粒層及外網識 1 * 層）分離出該光受《層。如第7圖中所示的，當接近該光受 髏層54時，該振動切片機之平台向被向前推進而獲得一約 厚50-300ιημ之切片。此切片之厚度必須夠厚而能將光受饅 切下並形成一由一層的光受體細胞所構成並有一薄明膠基 質58貼附其上之層狀物62。如第8圖中所示者，該切片62 沿該虛線被垂直地切下以生成一層狀物63。 如第9圖中所示者，該層狀物63接而被置放在一由豎 板66所形成之平板64上。帶有被置放在豎板66之間的層狀 物63之板64隨而被注入以融化的15-20¾明膠溶液俾以環繞 及包覆該光受體層54，而該明膠基質58亦為融化的明膠所 環繞及覆葦。如第10圖中所示者，一平坦的蓋板68被放在 豎板66之上以除去任何過量之融化的明膠並確立該移植物-之明確的厚度。該豎板66之高度可予以調節以製備具不同 厚度之移植物。 將所形成的容器67 (由被豎板66所分隔之兩個平板64 與68所構成且其圍繞一帶有被包埋在其内的光受體層54之 明膠板69)予以冷卻至室溫以導致該融化的明膠膠凝並形 一 14 - 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ297公釐） (請先閲讀背面之注意事項再填寫本頁) 524681 經濟部中央標準局員工消費合作社印製 Α7 Β7 五、發明説明（ll) 成一包封該光受體層54之載體薄片70。該光受體層54之外 節段（（未示出）面朝向該載體薄片70之一側71。 如第11圖中所示者，在容許該融化的明膠膠凝後，藉 由從豎板66將平板側向地滑移而小心地移除該層狀物之頂 端蓋板68俥以避免該明膠載體薄片70與該光受體層54之撕 裂。同樣地將該豎板66移除而暴露出該載體薄片70。為進 一步降低該明膠載體薄片70在該頂端蓋板68之移除時遭到 撕裂之危險，該頂端蓋板可被包覆以鐵弗龍薄膜（未示出） ，如此該.頂端蓋板之底部表面具有一固定於其上之平滑 的薄膜層。該頂端蓋板之移除係藉由解開位在該頂端蓋板 之上表面上的薄膜並從該豎板66舉起該平板。接而小心地 從該明膠載體薄片70剝去該鐵弗龍薄膜而將之移除。在一 解剖溶液（諸如一水性溶液）内之浸潤可加速剝除。 垂直切割該載體薄片70之相反兩端使成一供移植之適 當尺寸。如第13圖中所示者，該載體薄片的相反側72可被 削切一以相對於明膠板之頂端與底部表面呈鈍角及銳角來 切割--俾以形成一具有大約平行之側邊的移植物74。該移 植物74其琴削切之側邊72促進該移植物之一側邊72相對於 另一邊順利地滑移。該移植物74之表面應具有一大於Inna2 更佳為大於4ππη2之表面面積，或可大至在一外科儀器之操 作掌控範圍之内者，該外科儀器偽供經由一位於一宿主眼 内之的切口來移植該移植物。經此構建，該移植物74可包 腋一相當範圍的視網膜下表面。 欲製備供插入眼内用的移植物，從該平板64移出該移 一 15 - 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X297公釐） —---------裝-- (請先閱讀背面之注意事項再填寫本頁) 訂 線 524681 A7 B7__ 五、發明説明（12) (請先閱讀背面之注意事項再>寫本頁) 植物74 (第14圖）並令其形成一具有重疊側邊72與盤曲77之 渦形物76(第15圖）。該渦形物76之盤曲77彼此不影響另一 者這樣使得該等盤曲在隨後的解盤時不會阻礙該渦形物。 雖然其非目前所較偏好的，該渦形物76之盤曲77無必要重 疊；該移植物74之任一經盤捲的構形，因而該渦形物之直 徑較諸未盤捲的、薄片般形狀之移植物的側邊72之間的距 離要小以及因而光受體層54不受損害者，可依據本發明來 製備。 該移植物74 (包含有如上述之經切片與整平的視網膜 組織54以及該載體薄片70)之厚度傺僅大約的且隨供給者 材料之變動而變。此外，切片可予以加快而且振動切片機 厚度進一步地可從視網膜厚度之組織學測量而予以校準， 因而提對切片深度供進一步的導引。適當的切片厚度.或深 度可進一步琴由顯微鏡撿査和切片的觀察予以測定之。 經濟部中央標準局員工消費合作社印製 該明膠載體薄片70對該容易受損的光受體層54增加機 械強度與穩定性。結果，該經整平的視纲膜組織54即較不 容易受損害且更容易在移植期間内被操作之。明膠由於其 可撓性、可彎性、在體溫下會溶解之能力以及在溶解時對 神經組織明顯地不具毒性，而成為目前作為一包封劑之較 佳材料。但是，也可取代以亦具有明膠之所欲待徵的其他 組成，例如瓊膠或瓊脂糖。明顯地，未有！現到明膠會干 擾組織生長或介於該移植物74與下層的視網膜色素上皮之 間的後-移植相互作用。目前以明膠作為將該視網膜組織5 4層放在該包封物之内的黏著劑是較為適宜的。但是，其 -16 - · 本紙張尺度適用中國國家標準（CNS )‘八4規格（210X297公釐） 524681 A7 B7 _ 五、發明説明（13) 他的組成，包括諸如伴刀豆珠蛋白A之凝集素、小麥胚穿 凝集素或其在曝於光會膠凝或分解且其亦具有明膠之所欲 持徵之光反應劑，也可用來取代作為黏著劑之用。 有利地，該明膠載體薄片70或其他的包封物可額外地 供作為一有関下列任一者的載體：諸如成纖維細胞生長因 子之營養因子、包括諸如環孢靈素A之免疫抑制劑在内之 藥理試劑、諸如德塞美沙松（dexamethasone)之抗發炎劑 1 . ♦ 、抗血管形成因子、抗神經膠質劑以及抗有絲分裂因子。 在該包封物的溶解之時，該因子或試劑即能有效地對周遭 組識産生所欲之作用。其劑量可藉由已建立之實驗技術來 作決定。該包封物可含有可生物降解的聚合物以作為有關 可被包含在該基質内的藥理性物質之缓慢釋放劑之用。 作為機械性（亦即切片機）切片之一另一選擇，葭供給 者視網膜50亦可被化學性地切片。持別地，已知諸如卡因 酸之神經毒劑與氧缺乏對位於所有的視纲膜層内之細胞偽 有毒的，惟光受體與穆勒（Muller)細胞例外。因此，若該 供給者視網膜50以一適當的神經毒劑予以處理，可分離出 光受體層&4。此技術具有可維持視網膜穆勒細胞（其對卡 因酸和氧缺乏係相當地不敏感）與光受體細胞54之益處。 由於已知穆勒細胞（在生化上與結構上）幫助維持光受體細 胞，穆勒細胞加上光受體細胞之分離會是有利的。 若有需要，該移植物74可包含視網膜色素上皮細胞。 由於RPE傜強力地貼合至視網膜，視網膜從一供給者眼之 機械性剝離一般都會造成RPE從該視網膜分離出並維持者 一 17 — 本纸張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） I 訂 線 (請先閲讀背面之注意事項再瑣寫本頁) 經濟部中央標準局員工消費合作社印製 524681 A7 B7_’ 五、發明説明（14) 與脈絡膜之阽合。但經由酵素技術（諸如載述於Mayerson et a 1 . , Investi. Ophthalmol . Vis. Sni.. 26: 1599. 1609, 1985)之使用，可從供給者眼分離出帶有貼合的RPE 之視網膜。任擇地，其包含有一RPE細胞單細胞層之植入 物的製備可藉由從供給者組織收獲RPE細胞並將該經收擭 的RPE細胞如同一完整的單細胞層置放至一無毒的可撓性 组成上，或藉由以一單細胞層之解離的RPE細胞播種該一 组成並容許該等細胞生長成一匯合層。該可撓性組成供作 該等RPE細胞於包封及移植期間内之一安定支撑物之用。 其包含有脈絡膜、布魯赫氏膜或一合成的布魯赫氏膜 (亦即厚度為卜5μ的膠原蛋白）之移植物可被製備出。該脈 絡膜從眼之鞏膜内櫬被撕離（有或無貼合的RPE)並藉由作 徑向切割而被整平之。該供給者脈絡膜可如前述有關.光受 體细胞者予以包封之，和/或結合以如前述所製備的光受 震層54而形成一包含有一光受體層與一脈絡膜之層狀物， 其被包封在一明膠基質與超基質之内。 參照第16-18圖，其顯示一外科儀器78以供用於生成 一渦形物76以及將該渦形物植入宿主眼之移植位置内。該 外科儀華78以及本案之方法特別被設計成供用於從一供給 者視網膜分離出一完整的細胞薄片並經由一切口將之移植 至一接受者視網膜，該切口較諸將其呈解盤的薄Η般形狀 之該視網膜74插入所需之切口要小，以及該儀器78與方法 之進一步特徵在於維持經移植的組織層之組織結構。 第16圖大略地顯示一將完整的細胞結搆74植入一眼内 一 18 一 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ：Ζ97公釐） ----------參-------、訂------0 (請先閲讀背面之注意事項再楨寫本頁) 經濟部中央標準局員工消費合作社印製 524681 經濟部中央標準局員工消費合作社印製 Α7 Β7 五、發明説明（15) 的視網膜與支持組織之間的儀器78之一具體例。該儀器78 可以能無菌處理的丙烯酸、玻璃或其他適當的材料來製造 。該儀器78包含有一管狀體90，其具一開放端92以供承接 通常為平面的細胞結構74，一錐形通道或漏斗94以供將該 平面結構74盤捲成一渦形物76，以及一管狀尖端96以供插 入宿主眼内。如圖所示曁如此處所詋明者，該儀器78約為 10-15cm長，此乃供在齧齒類或低等靈長類内製作一植入 物之適當長度。其被插入眼之切口一眼入口一内之狹窄的 管狀尖端96必須夠長以延伸進入眼内至接近視網膜與支撑 的視網膜下組織之間處。關於該儀器78其狹窄的管狀尖端 96 *可使用不同的長度，而此依所使用的方法與所施用的 接受者而定。 如第21圖中所示者，該儀器78可在從該管狀體90至該 漏斗之·轉換處的儀器内側表面上包含一斜坡99。該斜坡99 導引一側72位在該移植物74之另一側下方以形成一渦形物 76。在該儀器73的這個具體例中，該載體薄片70之側邊72 可被垂直地切割而非削切以供形成移植物74。該斜坡99藉 由不容許該側邊接觸另一者來防止該移植物74之歪皺，並 且可以發揮令該渦形物之排列呈一特定的定位方向之作用 ，亦即該渦形物之一邊緣可被維持在一恃定的持定的定位 方向内。 如此處所示與敘述者，該儀器78之内徑在其開放端92 處為大約5mni而在其管狀尖端96處則約為80〇μ。該管狀尖 端96之直徑必須被作成其大小容許一完整的、經盤捲的細 一 19 一 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ 297公釐1 (請先閎讀背面之注意事項再填寫本頁) -裝- 訂 524681 A7 B7 五、發明説明（16) 胞結構一亦即，一渦形物76 —之通經其内以供植入而不導 致該渦形物之盤曲77在一端上産生剪應力並因此可能地造 成被包埋在其内的光受體層54之損害。職是，可使用不同 的管直徑，此傺依接受者以及該移植物74之大小而定。 更甚者，在該漏斗内從該開放端92之直徑至該狹窄的 管狀尖端96之直徑之轉換不可太突兀以致使移植物74産生 歪皺。因此，漏斗之斜度係漸進的以容許該移植物74之受 » - · 控制的盤捲。 如第18圖中所示者，該儀器78之狹窄的管狀尖端96可 被斜切以促進該儀器至眼内之插入以及該管狀尖端至帶有 一最低限的挫傷之眼的視網膜下組織内之推進。更甚者， 如第20圖中所示者，當該移植物之一端從該管狀尖端被排 放出時，該管狀尖端96其經斜切的邊緣98促進該移植物74 之漸進的解盤。該管狀尖端96之邊緣98以被斜切成45°為 佳。如一般在100處所指示者。該儀器78之狹窄的管狀尖 端96從該管狀尖端之邊緣98至該漏斗94之最窄末端沿其縱 軸最好是彎曲的。該管狀尖端之曲率100促進該儀器78在 眼内之操作，特別是該儀器至一位在眼之彎曲的壁上、介 於視網膜32與支持組織84之間的位置處之操作。該管狀尖 端之曲率100的半徑將梘操作方法以及宿主眼之曲率半徑 而定。 該儀器78進一步包括一柱塞構件85以幫助該移植物通 經該狹窄的管狀尖端96。如第18圖中所示者，該柱塞構件 85最好是一被承接於該管狀體90之開放端92内的薄管狀柱 一 2 0 一 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 1 _ 訂I 線 (請先閱讀背面之注意事項再读寫本頁) 經濟部中央標準局員工消費合作社印製 524681 經濟部中央標準局員工消費合作社印製 Α7 Β7 五、發明説明（17) 塞86，於是該柱塞關於該管狀體逋經該漏斗94並進入該管 狀尖端96之相對的推進驅策該經盤捲的細、胞结構76通經該 管狀體之漏斗以及通經該儀器78之管狀尖端。為減少由該 柱塞86與該細胞結構之間的直接接鐲所引起的對脆弱的細 胞结構76之損害，該經盤捲的細胞結搆76藉由一在該柱塞 之插入前被插入該誉狀體90之開放端92内並由明膠泡沫102 或其他柔軟可壓縮的材料所作成之間隔子而免於與該柱塞 86之直接接觸。該明膠泡沫102被導引置放在該經盤捲的 細胞结構76之頂端上而因此保護該盤捲免於與該機械性柱 塞之直接接觸。明膠泡沫係令人滿意的，因為其係為半固 體的且無毒性的。該柱塞86從該管狀體90之開放端92突出 一足夠之距離因而使得該柱塞之突出端88能被操作，即令 該儀器78之管狀尖端96偽在眼内亦然。操作該儀器78之較 佳的方法是一旦其内帶有該經盤捲的細胞結構76之管狀尖 端96被適當地放置在眼之視網膜下區域80之内，操作該柱 塞86以從該儀器之管狀尖端96排放出該經盤捲的細胞結構 76。雖然該柱塞86對該渦形物76至眼内之排放提供最大的 控制，由於對該渦形物76的損害之增加的可能性，在操作 該柱塞時某些慎重必須予以蓮用。 任擇地，該柱塞構件85可包含有用來對該管狀體90之 内含物施加流體壓力之構件（未示出）。就此方面而言，該 管狀體之開放端92被連接至一處於壓力下之流體源。流體 可經由連接至該儀器78之開放端92的管線被選擇性地供應 以取代其内含物。該流體可為黏稠的（例如2¾的羧甲基纖 一 21 — 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X297公釐） (請先閱讀背面之注意事項再域寫本頁) •裝· 524681 經濟部中夬標準局員工消費合作社印製 A7 B7 五、發明説明（18) 維素）或不黏稠的。特別就後者而言，可希望令明膠泡沫 或某種相當柔軟的間隔子材料位於該管内來發揮作為一機 械性柱塞之作用以將流體自正波植入的細胞结構分開。如 先前所討論的，明膠泡沫由於其傺為半固體的且若其自該 儀器被排放出將會無害地溶解，其對渦形物之保護僳令人 滿意的。雖然使用流體壓力以作為該柱塞構件85顯著地降 低對該渦形物76的損害之可能性，其亦造成對該渦形物76 從該儀器78之排放的控制程度上一顯著的降低。 如在專利申請案序列编號07/566,996 (其在此被併入 本案以為參考文獻）中所示及敘述的，該儀器可予以加入 許多特徵以促進一持殊的外科。如第17圖中所示者，該儀 器可包含有一通長平行於該儀器78之管腔108。如此處所 用者，管腔108意指任一管狀容器，不論其傜為分開提供 的或是被形成如一位在該儀器78之外側上的通道。該管腔 108具有一通常臨接該儀器78之管狀尖端96並最好是相對 於該管狀尖端略微地提前之的末稍端110。該管腔108之基 部端112傜遠離該末稍端110且可被設置有一供與一處於壓 力下的流體連接之連接器。因此，該管腔108可從其末稍 端110排放一流體流而生成一在該儀器78之前的流體空間 。該儀器78之管狀尖端96通常根隨著由該流體打通之通道 因而使該儀器與眼組織之直接接觸減至最低程度。該管腔 108末稍端110可被斜切成45°以促進該儀器78之推進，持 別是當該流體未從該管腔被排放出之時候。該末稍端110 最好是被斜切成45° 。從該管腔108被排放出的流體可為 - 22 - 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 裝 訂 線 (請先閲讀背面之注意事項再瑣寫本頁) 經濟部中央標準局員工消費合作社印製 524681 A7 B7 五、發明説明（19) 一生理鹽液，或某些不會損害眼組織之其他流體。不同的 例子例如抗氧化劑、抗發炎劑、抗有絲分裂劑與局部麻醉 劑可被提供於該流體内以供眼或植入的組織之治療。 視外科之類型，該儀器亦可包含一光纖纖維（未示出） ，其大體上與該管腔108平行地延伸並被置放在該管腔與 該管狀體90之間。該光纖纖維以兩種方式來促進該儀器78 之操作以及該移植物74之適當的放置：一光源可被提供於 該光纖維之基部繞處於是該纖維在該儀器78之管狀尖端 96處提供光，俾以幫助通經瞳孔之視覺觀察程序；任擇地 ，一透鏡可被邊供於該光纖纖維之基部端處於是該纖維可 被用來供在該儀器之管狀尖端處的直接觀察。此外，該光 纖纖維可容許雷射光燒灼以控制視網膜下出血。 該儀器78可進一步包含一第二管腔（未示出），其大體 上與第一管腔108平行地延伸並被置放在該管腔1〇3與該管 狀體90之間。該第二管腔容許抽吸源自該儀器78之管狀尖 端96的物質。該管腔之基部端可被連接至一吸取源俾以移 除過量的流體與碎屑。 該儀器78可進一步包含一對導線（未示.出），其终止於 其末稍端之電極内。該電極容許血管之燒灼。該導線之基 部端可被連接至一電源以密封破裂的血管。亦可以將該導 線併入該儀器78之管狀體90的壁上。 當然，關於該等任擇的具體例所述之二或二種以上的 特徵，如在待定情況下有所需要，可將之組合在一起。 將一渦形物76移植至一眼之視網膜下區域的方法包括 一 2 3 - 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 訂 線 (請先閱讀背面之注意事項再梦寫本頁) 經濟部中央標準局員工消費合作社印製 524681 A7 _ B7___ 五、發明説明（20) 如前述將一可移植的物質（諸如視網膜色素上皮組織、脈 絡膜組織、布魯赫氏膜和/或視網膜細胞）組合成一移植物 74。將可瞭解到，該可移植的物質可被形成一不具明膠載 體薄片之移植物且仍落在本發明之範疇内。但較佳地，該 移植物與一載體薄片70被組合之。該移植方法提供將該移 植物74放置到該儀器内位於該管狀體90之開放端92處，而 該移植物74與該管狀體之内壁接合。該移植物74其一端73 首先被置放在該管狀‘體90之開放端92内，因而該載體70會 被盤捲，而使光受體層54之外節段面朝向所形成的渦形物 76之盤曲77之外側，因而該渦形物將於該視網膜下區喻内 解盤而使該光受體層之外節段面朝向宿主眼之色素上皮層 84。該儀器73之管狀尖端96被加帽以118且該管狀體90被 充填以黏彈性流體120，其促進該移植物至該錐形通道或 漏斗内之前進。該移植物滑動地進入該漏斗内接合漸進地 變窄之錐形表面，這造成該移植物逐漸地盤捲。當該漏斗 94之内壁夠窄時會造成該載體薄片70之倒邊72變成緊密的 ，該薄片70之一側72滑行至該載體薄片之另一側的下方， 此部分地傜由於該載體之經削切的側邊所致。當該側邊邊 緣變成緊密時，該等經削切的側邊72防止該載體薄片70之 任何歪皺。在一示於第21圖之任擇的具體例中，當該儀器 78之内壁夠窄時而使該移植物74之側邊72彼此相靠近時， 斜坡99導引一位在另一側邊之下方的側邊72開始該渦形物 76之盤捲。在該漏斗94内之某一點處，該盤捲物76之盤曲 77被充分地壓縮致使該年彈性流體120無法再經由該漏斗 一 24 — 本纸張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） I——^------參------1T-------^ (請先閲讀背面之注意事項再填寫本頁) ♦經濟部中央標準局員工消費合作社印製 524681 A7 B7 五、發明説明（21) 強迫該盤捲物。一明膠泡沫間隔子102被置放在該盤捲物 76之頂端，一球狀體104被置放在該儀.器之開放端92的頂 端上以生成一真空，於是該流體120與該渦形物留存在該 儀器78内。一注射針106可經由該球狀體104被插入以注入 更多之如所需要的流體120。該柱塞86經由該球狀體104被 插入該管狀體90之開放端92内並被操作成與該明膠泡沫間 隔子102相接觸。該柱塞86被小心地推進以強迫該移植物 4 I ， 74經由該漏斗94更進一步地將該移植物盤捲成一渦形物76 並進入及通經該管狀尖端96。 好好準備宿主眼俾使其減少出血及外科挫傷。一至視 網膜下間隙之鞏膜睫狀體垣部途徑（第20圖）係較佳的，但 亦可使用其他的途徑，例如跨越角膜及鞏膜者。在睫狀體 垣部作一小切口，其大小足容插入外科儀器78(約0.75min-約2.0mm)。在玻璃體切除後，經由一第二個睫狀體垣部入 口注入經冷卻之平衡的鹽水溶液至眼112之玻璃體室内來 冷卽該眼，俾以避免該渦形物76之載體薄片70在外科處理 過程中之解離。藉由在視網膜内作一切口並將平衡的鹽水 溶液注入視網膜下區域以在視網膜82的移植位置處形成一 小泡80，而將位在移植位置處之一部分的視網膜82從色素 上皮細胞内襯δ4予以掀離。若該儀器78包含一管腔108， 該視網膜藉由一從該管腔排放出的灌流液（諸如生理鹽液 般流體、羧甲基纖維素或1-2¾琉璃酸）的溫合作用力而被 剝離俾以生成一小泡80。有利地，該流體可額外地含有抗 氧化劑、抗發炎劑、麻醉劑或其會降低宿主視網膜82之代 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X 297公釐） — 訂 線 (請先閲讀背面之注意事項再續寫本頁) 經濟部中央標準局員工消費合作社印製 524681 A7 B7 五、發明説明（22) 謝需求的試劑。 在其管狀尖端96處帶有該渦形物76之儀器73經由該睫 狀體垣部入口，該玻璃體室而被插入視網膜下間隙。如第 20圖所例示的，接而操作該儀器78致使該管狀尖端96之邊 緣與該小泡80之切口 122成一直線。該儀器78之整個頂尖 被插入該小泡80内，而該渦形物76藉由小心地推進該柱塞 而被排放出。該渦形物76從該管狀尖端96之斜切邊緣98被 1 -" 排放出並在其固有的解盤記億下，於其從該斜切邊緣被排 出時解開盤捲，於是光受體層54之外節段即面朝向該色素 上皮層84。若該渦形物76没有完全地解盤，可使用微牙籤 將該移植物74完全地解盤。 該小泡80接而藉由該小泡内流體之疏散或藉由tenipa-nade被抽氣之，致使該移植物74呈夾層式排列被置於靠近 視網膜82與色素上皮内襯84之所欲位置處。該小泡80之切 口 122可被燒灼地閉合。該明膠載體薄片70—旦接近正常 體溫即會溶解。該鞏膜切口之邊緣在移走鑷子後被接連並 使用標準眼科方法縫合之。 如在母案申請案序列编號07/566,996中所示及敘述的 ，一跨越脈絡膜、鞏膜及角膜之外科方法可被用來作為上 述睫狀體垣部方法之一任擇的實施例。除了進入之點外， 該外科技術大體上同於上述之方法。 •基於上述，可看到本發明之數個目的之達成以及其他 益處之獲得。 由於上述外科儀器、物質組成與方法之各種不同的變 、 - 26 - 本紙張尺度適用中國國家標準（CNS ) A4規格（210x297公釐） f (請先閱讀背面之注意事項存攻寫本頁) -装· 訂 524681 A7 B7 五、發明説明（23) 化在不逸脫本發明之範疇下可被做出，則包含在上述敘述 内或顯示於隨文所附圖式内之所有事務將被視為偽用來解 說本發明而非用以為限制本發明。 (請先閲讀背面之注意事項再妒寫本頁) 經濟部中央標準局員工消費合作社印製 7 2 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐）524681 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs Α7 Β7 V. Description of the Invention (1) Background of the Invention This application was filed in 1993. 3.16 and 19908. 13. Co-pending copending applicaton with serial number 0. 8/033, 105 and 07 / 566,996, part of the continuation-in-part application, which was re-used as 19δ9 · δ · 14 · Part of the continuation application. The present invention is generally broader than surgical instruments and techniques. More specifically * Indeed, the present invention is directed to a surgical tool for transplanting thin, epithelial and / or choroidal tissue of retinal cells in a volute configuration through a standard-sized incision in the eye. A graft for transplantation into the subretinal area of the eye and a method for reconstructing a malnourished retina, retinal pigment epithelium, and choroid. The retinal ridge is a sensory epithelial surface that lines the posterior chamber of the eye, receives an image formed by the lens, converts this image into a neural pulse, and transmits this information to the brain through the optic nerve. The retina contains several layers, namely the ganglion cell layer, the inner reticular layer, the inner granule layer, the outer reticular layer, the outer granular layer, the inner and outer segments of the photoreceptor. The outer granule layer contains a cell body of photoreceptor cells with the inner and outer segments as extensions of the cell body. The choroid is a pseudo-vascular membrane that contains a large number of branched pigment cells between the retina and the scleral coat of vertebrate eyes. Located directly between the choroid and the retina is the retinal pigment epithelium, which forms a close structural and functional relationship with the photoreceptor cells. Several types of blindness are mainly related to defects in the retina, retinal pigment epithelium, choroid, or other possible factors (such as glare, vision 5 — this paper size applies Chinese National Standard (CNS) A4 specification (210 × 297 (Mm) —-- ί ------ install-- (please read the notes on the back before filling in this page) Order printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 524681 A7 B7 V. Description of Invention (2) Omentum detachment, intraocular hemorrhage) caused by loss of photoreceptor cells. In several retinal degenerative diseases, selected cell populations are lost. In particular, in plaque degeneration and pigmented retinitis, its retinal photoreceptors are degraded, while other cells located in the retina and the central junction of the retina are maintained. In an effort to restore a damaged retina that was previously considered to be irreparable, researchers have suggested various types of transfer materials and transplantation techniques, none of which constitute an effective type to reconstruct an undernourished Retina. The transplantation of retinal cells into the eye can be traced to Royo et al. Growt k21: 313-336 (1959), in which the embryonic retina is transplanted into the anterior chamber of the mother's eye. Various cells, including photoreceptors, have been reported to live. Del Cerro can then repeat and extend these experiments (del Cerro e t a 1. , Invest. Ophthalmol. Vis. Sci. 26 «1182-1185, 1985). Immediately after Turner et al. , Dev. Brain Res,. 26: 91-104 (1986) showed that neonatal retinal tissue can be transplanted into the retina. In a related study, Simmons et al. , Soc. Meurosc i. Abstr, Iz: 668 (1984) demonstrated that embryonic retinas can be transplanted intracranially and survived, showing fairly normal development, being able to innervate central structures, and activating these structures in a light-dependent manner. What's more, these intracranial grafts can stimulate light-dependent behavioral responses (pupilary L reflexes) regulated by the host's nervous system 0Klassen et a 1. , Exp. Neurol, 102: 102-108 (1988) and Klassen et al., Proc. Na11 > AnMU. USA 84: 6958-6960 (1987). 6-This paper size applies to China National Standard (CNS) A4 (210X297 mm) — gutter (please read the precautions on the back before filling this page) 524681 Printed by A7 B7 of the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (3) L ί and Turner, Exp. Eye Res, 47 ♦ 911 (198 &) 曽 Suppose that the transplantation of retinal pigment epithelium (RPE) into the lower retinal space is used to replace the defective with healthy wild-type related controls in RCS malnourished rats Of mutant RPE cells as a treatment modality. According to their method, RPE was isolated from 6 to 8 days old black-eyed rats and transplanted into the subretinal space using a paradigm of penetrating the sclera and choroid. An RPE injection (40,000-60,000 cells) of lnil was placed into the subretinal space using a 10 ml injection needle followed by a 30-gauge needle at the incision site. However, this method destroys the cellular polarity and natural tissue structure of the host retinal pigment epithelium required for the graft. del Cerro (del Cerro et a 1. , Invest. Ophthalmol. Vi ^ W. 21 _... 11δ2-1185, 1985) reports a method for transplanting a tissue slice into the anterior chamber or host retina. These bright slices were obtained by cutting the neural retina from the donor's eye. The retina was then cut into appropriate tissue strips, which were then injected into the appropriate location using a 30 gauge needle or emblem pipette, and the width of the strip was limited to the inner diameter of the needle (25 〇μπ) and the length is less than lnun. Although del Cerro reports viable intraocular transplantation of the omentum strip, it has noted certain limitations to this approach. For example, his technology cannot tolerate the replacement of cells that are losing (such as photoreceptors), but always include a mixture of retinal cells. Therefore, with such a graft, it is impossible to properly reconstruct the malnourished retina lacking a fixed cell population (such as photoreceptors). del Cerro e t a 1. , Neurosc i. Lett. 92 »21-26, January 7 — This paper size applies to the Chinese National Standard (CNS) A4 (210X297 mm) I binding line (please read the precautions on the back before filling this page) 524681 Employees of the Central Standards Bureau of the Ministry of Economic Affairs Cooperative printed A7 B7 V. Description of the invention (4) 1988 Re-Report on a method for transplanting dissociated neural retinal cells. In this method, the donor retina is cut into small pieces, incubated in trypsin for 15 minutes, and developed into a single cell suspension by aspirating through a finely drawn pipette. In contrast to the Li and Turner method discussed above, this method destroys the organized, native structure of the graft, including the donor's outer granule layer. The strict fibrillar structure of the photoreceptor (with the segments towards the outer part of the pigmented epithelium and the end of the processus facing the outer reticulum) is lost. Furthermore, no method has been used to isolate and purify any given retinal cell population (such as photoreceptors) from other retina cells. The inventor of this case believes that in order to restore a reasonable degree of vision, it is necessary to maintain the photoreceptors in an organized outer granular layer structure. This conclusion is based on the optical characteristics of the known photoreceptors (the outer segment acts as a light guide) and clinical evidence (which shows that intra-retinal geometric folds or similar (even very small) ruptures can be severe Degradation of vision. Invention To explain, therefore, among the objects of the present invention, it is noted that there is a method for providing a considerable amount of tissue harvested from the eyes of a supplier; providing such a method, a considerable amount of which The harvested tissue is shaped so that the harvested tissue can be inserted into a standard large incision in the eye; a method is provided in which the polarity and tissue structure of the cells are harvested Maintaining in the graft; and providing a method for transplanting the graft into the subretinal region of one eye. Further, among the several purposes and characteristics of the present invention, it may be noted that the paper size is applicable to China National Standard (CNS) A4 specification (21〇X: 297mm) l · --r ------ install-- (please read the precautions on the back before filling this page), π line _ 524681 A7 B7 V. Description of the invention (5) To one is to provide a graft for the reconstruction of a malnourished retina or to rescue an endogenous photoreceptor cell of an individual suffering from an epidemic or acquired human retinal disease. Illness which causes gradual loss of vision protection and, in turn, eventually causes malnutrition, dysfunction and / or loss of the cone; Such a graft is provided which promotes the regrowth of photoreceptor axons by maintaining the polar structure of the photoreceptor at the time of transplantation and its close proximity to the nearby postsynaptic target. Among other things, it is noted that one of the purposes and features of the present invention is to provide a surgical tool for use in the implantation method, which forms the graft for insertion into a large-sized incision; and A surgical tool for transplantation that allows proper localized placement of the retina and protection of photoreceptors, retinal pigment epithelium, choroidal tissue, and / or Bruch's membrane at the surgical device Avoid damage before and when you put it in your eyes. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs ---- ί -------- Loading-(Please read the precautions on the back before filling out this page). In general, the implantation method includes coiling An implantable substance that has a thin sheet-like shape that can form a volute. The coils of the volutes do not affect each other, so that the subsequent unwrapping of the implantable substance is substantially restored to its original sheet-like shape. An incision is made in the host eye for inserting a vortex at a location between the host's retina and the underlying tissue. The cut is smaller than the cut required to insert the implantable substance in the form of a thin sheet-like shape. One end of the scroll is first inserted into the host's eye through the incision to a position between the retina and the underlying tissue. After the scroll is inserted, the disc is laid flat in a sheet-like shape. Between the retina of the host eye and the underlying tissue and the incision is closed. This paper size applies to China National Standard (CNS) A4 (21 × 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 524681 A7 __B7 V. Description of the invention (6) Generally speaking, the graft for transplantation contains There is a layer of non-toxic, flexible composition that will generally dissolve at body temperature and a substance to be transplanted that is coiled to form a volute. One end of the scroll can first be inserted into a place for transplantation through an incision (the size of which depends on the cross-sectional area of the scroll), and then the unwrapped disk lies flat on the sheet. The location of the transplant is ϋ. Generally, the implant used to form a volute includes a tubular body that is open at one end and has a funnel. A carrier first enters from the open end of the tubular body and feeds through the funnel along its guard. The engagement of the carrier with the inner surface of the funnel as it is fed through the funnel causes the carrier to coil into a volute. The presence of the scroll starts at one narrow end of the funnel. Other objects of the present invention are partly known, and part of them will be specified hereinafter. Figure 槪 to explain Figure 1 is a frozen section photo of one of the normal rat retina as described in Example 1; Figure 2. It is a photograph of a frozen section of one rat's retina that becomes blind after continuous illumination as described in Example 1. Figure 3 is a schematic diagram of a donor retina; Figure 4 is a flattened retina- Schematic diagram; Figure 5 is a schematic diagram of a flattened retina placed on a substrate; Figure 6 is a sliced retina placed on a substrate-10-paper size Applicable to China National Standard (CNS) Α4 specification (210 X 297 mm) I --- r ------ clothing ------, 玎 ------ ^ (Please read the Please fill in this page again for attention) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 524681 Μ B7__ 5. Description of the invention (7) Schematic diagram; A schematic view of one of the layers; FIG. 8 is a schematic plan view of the top of the layer of FIG. 7, which shows a graft (dashed line) including a photoreceptor cell layer and a supporting and stabilizing matrix; FIG. 9 The picture shows a layer placed on a flat plate formed by spacers A schematic diagram; FIG. 10 is a schematic view of one layer is disposed on the gelatin was added coverslips plate is injected;. Figure 11 is a schematic diagram of the top plate being slid away from the ground; Figure 12 is a schematic diagram of one of the formed grafts; Figure 13 is a schematic diagram of a graft being cut; Figure 14 is one of the grafts being removed from the plate for transplantation Figure 15 is a perspective view of a scroll; Figure 16 is a front view of an instrument for coiling and implantation of the graft: and the coiled graft is located in the funnel of the instrument Figure 17 is a side view of one of the instruments and a lumen is attached to the outside of the instrument; Figure 18 is a side view of the instrument and a plunger that advances the scroll; Figure 19 It is a horizontal cross-sectional view through one eye, which illustrates a surgical method using the instrument to partially extend across the eye;-11-^ 's scale is applicable to China National Standard (CNS) A4 size (210 X 297 mm) Button sewing thread (please read the precautions on the back before filling out this page) 524681 A 7 B7 V. Description of the invention (0 Figure 20 is a horizontal cross-sectional view through one glance, which illustrates an example of using the instrument inserted into a bubble Surgical method of ciliary body; Figure 21 is a horizontal cross-sectional view taken along line 21-22 of Figure 16 and illustrates a slope located within the instrument. Relevant reference signs in the figures and figures of the drawings mean related parts. Xue Ming specified that as used herein, the term "provider" pseudo-refers to the same or different organisms relative to the host, and the term "provider organization" refers to the same or relative Tissues obtained by harvesting or cultured tissues obtained by foreign organisms. Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page). Several types of blindness (such as pigmented retinitis, retinal detachment, plaque degeneration, and blindness related to light exposure) It is the loss of light protection in the eyes. However, the destruction of photoreceptors does not necessarily result in the loss of the remaining retina or axons that connect the retina to the brain. Surprisingly, the inventors have found that by replacing damaged photoreceptors with photoreceptors harvested from a supplier and maintained at their original tissue structure and cell polarity, to some extent Vision is recoverable. What's more, as further described in Commonly Examined Patent Application Serial No. 08/033, 105 (which is incorporated herein as a reference), it is harvested from the light receptor of a supplier's eye Rod transplantation can `` rescue endogenous cone light receptors located within the retina and therefore restore or preserve the visual acuity of cone light receptors that still exist. That is, the present inventors have discovered that the transplanted rods exert a nutritional effect on the endogenous cone. Figure 1 is a photograph of a frozen section of a normal rat's retina. 12 — This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm). Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 524681 A7 B7___ 5. Description of the invention (9). Figure 2 is a photograph of a frozen section of a rat's retina after continuous illumination [breaking the photoreceptor (exolemma) layer but leaving most of the other retinal layers and cells intact]. In these and subsequent drawings, the retina or its layers, such as the ganglion cell layer (" G "), the inner network recognition layer (" IPL"), the inner granular layer ("INL"), the outer mesh Layer ("〇PL"), outer granule layer ("ONL"), inner segment (" IS "), outer segment (" 〇s "), and retinal pigment epithelium (" R PE ") from top to bottom The photoreceptor graft for implantation through all the mouths is prepared according to a method of the present invention with reference to FIG. 3, and the thickness of the incision is thinner than that of the graft. It is small. However, the graft may contain other implantable substances, such as other retinal cells, antiviral and antibiotics, and / or other pharmacological indicators. By removing a donor from the eye of a donor The retinal membrane 50 (including the inner retinal membrane layer 52 and the photoreceptor layer 54) is used to prepare a * graft containing photoreceptor cells. The number is calculated from the position near the center of the retina to its outer edge The second resection (see Fig. 8) is performed to flatten the donor retina 50 (Fig. 4). If necessary, the cutting can be performed in other directions. As shown in FIG. 5, the flattened retina 56 is placed on a gel plate 58 with its light-receiving side 54 facing down. The surface has been flattened to provide a flat surface 60 parallel to the blade of a vibrating microtome. The gelatin plate 58 is fixed to a conventional chuck of the vibrating microtome. 4-5% to melt A gelatin solution is deposited at the interface 61 adjacent to the flattened retina / gelatin surface, and the gelatin is attracted by capillary action under the flattened retina, causing the flattened retina to float on the gelatin plate 58. Xunyi 13 — This paper is again applicable to China National Standard (CNS) A4 specification (21 × 297mm) I Binding line (please read the precautions on the back before filling this page) 524681 A7 B7 Official Director of the Ministry of Economic Affairs Printed by Fei Cooperative 5. Description of the invention (10) The excess melted gelatin was quickly removed, and the floating, flattened retina was subsequently removed with ice cold Ringer's solution (R inerts sol lit ion). Cool to about 4 ° C, this Ringer's solution The gelatin block is wound to cause melting gelatin to gel. The flattened retina 56 is thus attached to the gelatin block. As shown in FIG. 6, the inner retinal portion 52 is applied from the top downward by about 20 to 50 μm. The slice is close to the photoreceptor layer 54, and the light receiving layer is separated from the inner layer of the retina (ie, the ganglion cell layer, the inner reticulum layer, the inner granule layer, and the outer retina 1 * layer). As shown in Figure 7, when approaching the light-receiving layer 54, the platform of the vibrating slicer is pushed forward to obtain a slice of about 50-300 μm thick. The thickness of this slice must be thick enough to light The anchor is cut out to form a layer 62 composed of a layer of photoreceptor cells with a thin gelatin matrix 58 attached thereto. As shown in FIG. 8, the slice 62 is cut vertically along the dotted line to generate a layer 63. As shown in Fig. 9, the layer 63 is then placed on a flat plate 64 formed by a vertical plate 66. The plate 64 with the layer 63 placed between the vertical plates 66 is then injected with a melted 15-20 ¾ gelatin solution to surround and cover the photoreceptor layer 54, and the gelatin matrix 58 is also It is surrounded and covered by melting gelatin. As shown in Figure 10, a flat cover plate 68 is placed on the riser 66 to remove any excess melted gelatin and establish the clear thickness of the graft. The height of the riser 66 can be adjusted to prepare grafts having different thicknesses. The formed container 67 (consisting of two flat plates 64 and 68 separated by a riser 66 and surrounding a gelatin plate 69 with a light-receiving layer 54 embedded therein) was cooled to room temperature This melted gelatin gelled and formed a shape 14-This paper size applies to the Chinese National Standard (CNS) A4 specification (210 × 297 mm) (Please read the precautions on the back before filling this page) 524681 Employees of the Central Standards Bureau of the Ministry of Economic Affairs Printed by the consumer cooperative A7 B7 5. Description of the invention (11) A carrier sheet 70 encapsulating the photoreceptor layer 54 is formed. The outer segment (not shown) surface of the photoreceptor layer 54 faces one side 71 of the carrier sheet 70. As shown in FIG. 11, after allowing the melted gelatin to gel, The plate 66 slides the plate laterally and carefully removes the top cover 68 of the layer to avoid tearing of the gelatin carrier sheet 70 and the photoreceptor layer 54. Similarly, the vertical plate 66 is moved In addition, the carrier sheet 70 is exposed. To further reduce the risk of the gelatin carrier sheet 70 being torn when the top cover 68 is removed, the top cover may be covered with a Teflon film (not shown) Out), so it should be. The bottom surface of the top cover plate has a smooth film layer fixed thereon. The top cover is removed by releasing the film on the upper surface of the top cover and lifting the flat plate from the vertical plate 66. The Teflon film was then carefully removed from the gelatin carrier sheet 70 and removed. Infiltration in an anatomical solution, such as an aqueous solution, can speed the removal. The opposite ends of the carrier sheet 70 are cut vertically to an appropriate size for transplantation. As shown in FIG. 13, the opposite side 72 of the carrier sheet may be cut to make an obtuse and acute angle with respect to the top and bottom surfaces of the gelatin sheet-- 俾 to form a side having approximately parallel sides Graft 74. The cut sides 74 of the moving plant 74 facilitate smooth sliding of one side 72 of the graft relative to the other. The surface of the graft 74 should have a surface area larger than Inna2, more preferably 4ππη2, or it can be as large as within the control range of the operation of a surgical instrument, which is supposed to pass through a surgical instrument located in the eye of a host An incision is made to transplant the graft. With this construction, the graft 74 can cover a considerable area of the subretinal surface of the axilla. To prepare an implant for insertion into the eye, remove it from the tablet 64 and remove it 15-This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) —--------- pack- -(Please read the precautions on the back before filling this page) 524681 A7 B7__ V. Description of the invention (12) (Please read the precautions on the back before writing this page) Plant 74 (Figure 14) and order It forms a volute 76 with overlapping sides 72 and a convolute 77 (Figure 15). The coils 77 of the volutes 76 do not affect each other so that the coils do not hinder the volutes during subsequent unwinding. Although it is not currently preferred, the coils 77 of the scroll 76 need not overlap; any of the coiled configurations of the graft 74, therefore, the diameter of the scroll is greater than that of uncoiled, thin slices. A generally shaped implant having a small distance between the sides 72 and thus the photoreceptor layer 54 is not damaged can be prepared according to the present invention. The thickness of the graft 74 (containing the sliced and flattened retinal tissue 54 and the carrier sheet 70 as described above) is only approximately and varies with the material of the donor. In addition, the sectioning can be accelerated and the thickness of the vibrating microtome can be further calibrated from the histological measurement of the thickness of the retina, thus providing further guidance on sectioning depth. Proper section thickness. Or the depth can be further measured by microscopic inspection and observation of section. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs The gelatin carrier sheet 70 adds mechanical strength and stability to the photoreceptor layer 54 which is easily damaged. As a result, the flattened retinal membrane tissue 54 is less susceptible to damage and is more easily manipulated during transplantation. Because of its flexibility, bendability, ability to dissolve at body temperature, and apparently non-toxicity to nerve tissue when dissolved, gelatin has become a better material as an encapsulant. However, it is also possible to substitute other components which also have gelatin as desired, such as agar or agarose. Obviously, none! It has now been shown that gelatin may interfere with tissue growth or a post-graft interaction between the graft 74 and the underlying retinal pigment epithelium. Gelatin is currently suitable as an adhesive for placing the 54 layers of retinal tissue within the encapsulant. However, its -16-· This paper size applies to the Chinese National Standard (CNS) '84 size (210X297 mm) 524681 A7 B7 _ V. Description of the invention (13) His composition, including such as concanavalin A Lectins, wheat germ penetrating lectins, or photoreactive agents that gel or decompose when exposed to light and also have the desired characteristics of gelatin can also be used instead of adhesives. Advantageously, the gelatin carrier sheet 70 or other encapsulant may additionally be used as a carrier for any of the following: trophic factors such as fibroblast growth factors, including immunosuppressants such as cyclosporin A Included pharmacological agents, anti-inflammatory agents such as dexamethasone 1. ♦ Anti-angiogenic factors, anti-glia and anti-mitotic factors. When the encapsulate is dissolved, the factor or agent can effectively produce the desired effect on the surrounding tissue. The dosage can be determined by established experimental techniques. The encapsulant may contain a biodegradable polymer for use as a slow release agent for a pharmacological substance that can be contained in the matrix. As an alternative to mechanical (i.e., microtome) sectioning, the donor's retina 50 can also be chemically sectioned. In particular, neurotoxins such as caine acid and oxygen deficiency are known to be pseudotoxic to cells located in all retinal membrane layers, with the exception of photoreceptors and Muller cells. Therefore, if the donor retina 50 is treated with an appropriate nerve agent, the photoreceptor layer & 4 can be separated. This technique has the benefit of maintaining retinal Muller cells, which are relatively insensitive to cainic acid and oxygen deficient lines, and photoreceptor cells54. Since Mueller cells are known (both biochemically and structurally) to help maintain photoreceptor cells, the separation of Mueller cells plus photoreceptor cells would be advantageous. If desired, the graft 74 may include retinal pigment epithelial cells. Because RPE fits tightly to the retina, mechanical peeling of the retina from a supplier's eye will generally cause RPE to separate from the retina and maintain it. 17 — This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 (Mm) I Thread (Please read the notes on the back before writing this page) Printed by the Consumers 'Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 524681 A7 B7_' V. Description of the invention (14) Combination with choroid. But via enzyme technology (such as described in Mayerson et a 1. , Investi. Ophthalmol. Vis. Sni. . 26: 1599. 1609, 1985), the retina with a conformable RPE can be isolated from the donor's eye. Optionally, an implant comprising a single cell layer of RPE cells can be prepared by harvesting RPE cells from a donor tissue and placing the collected RPE cells as a complete single cell layer into a non-toxic Flexible composition, or by seeding the composition with dissociated RPE cells in a single cell layer and allowing the cells to grow into a confluent layer. The flexible composition is used as a stable support for the RPE cells during the encapsulation and transplantation period. Grafts containing choroids, Bruch's membranes, or a synthetic Bruch's membrane (that is, collagen with a thickness of 5μ) can be prepared. The choroid is torn from the scleral condyle of the eye (with or without conforming RPE) and flattened by radial cutting. The donor choroid can be related as previously described. The photoreceptor cells are encapsulated and / or combined with the light shock layer 54 prepared as described above to form a layer including a photoreceptor layer and a choroid, which is encapsulated in a gelatin matrix With supermatrix. Referring to Figures 16-18, a surgical instrument 78 is shown for generating a volute 76 and implanting the vortex into a transplantation site in a host eye. The surgical instrument Hua 78 and the method of the present case are specifically designed for separating a complete cell sheet from a donor retina and transplanting it through a mouth to a recipient retina. The thin ridge-like shape of the retina 74 requires small incisions for insertion, and the instrument 78 and method are further characterized by maintaining the tissue structure of the transplanted tissue layer. Figure 16 roughly shows a complete cell structure 74 implanted into an eye 18 18 a paper size applies the Chinese National Standard (CNS) A4 specification (210 ×: Z97 mm) ---------- ref. ------- 、 Order ------ 0 (Please read the notes on the back before writing this page) Printed by the Consumers 'Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 524681 Printed by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs Preparation A7 B7 V. Description of the invention (15) One specific example of the instrument 78 between the retina and the supporting tissue. The instrument 78 may be made of acrylic, glass or other suitable materials that can be aseptically processed. The instrument 78 includes a tubular body 90 having an open end 92 for receiving a generally planar cell structure 74, a conical channel or funnel 94 for coiling the planar structure 74 into a volute 76, and a A tubular tip 96 is provided for insertion into the host's eye. As shown in the figure, as shown here, the instrument 78 is about 10-15 cm long, which is a suitable length for making an implant in rodents or lower primates. The narrow tubular tip 96 that is inserted into the incision, the entrance, and the eye of the eye must be long enough to extend into the eye to between the retina and the supporting subretinal tissue. Regarding the instrument 78, its narrow tubular tip 96 * can be used in different lengths, depending on the method used and the recipient to be administered. As shown in Figure 21, the instrument 78 may include a ramp 99 on the inside surface of the instrument from the tubular body 90 to the transition of the funnel. The slope 99 guides one side 72 below the other side of the graft 74 to form a volute 76. In this specific example of the instrument 73, the side edges 72 of the carrier sheet 70 may be cut vertically instead of being cut for forming the graft 74. The slope 99 prevents the wrinkle of the graft 74 by not allowing the side to contact the other, and can play a role in making the arrangement of the scrolls a specific positioning direction, that is, the scrolls An edge can be maintained in a fixed, fixed orientation. As shown here and the narrator, the inner diameter of the instrument 78 is approximately 5 mni at its open end 92 and approximately 80 μm at its tubular tip 96. The diameter of the tubular tip 96 must be made to a size that permits a complete, rolled thin one. 19 This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 × 297 mm 1) (please read the note on the back first) (Please fill in this page again for more information)-Binding-Order 524681 A7 B7 V. Description of the invention (16) Cell structure one, that is, a volute 76-passes through it for implantation without causing the volute's disk The bend 77 produces a shear stress on one end and may therefore cause damage to the photoreceptor layer 54 embedded therein. It is possible to use different tube diameters, depending on the recipient and the size of the graft 74 What's more, the conversion from the diameter of the open end 92 to the diameter of the narrow tubular tip 96 in the funnel should not be too abrupt to cause the graft 74 to wrinkle. Therefore, the slope of the funnel is progressive To allow coils of the implant 74 to be controlled by »-·. As shown in Figure 18, the narrow tubular tip 96 of the instrument 78 can be beveled to facilitate insertion of the instrument into the eye and the tube Tip to retina with a minimally contused eye Advancement in the lower tissue. Furthermore, as shown in Figure 20, when one end of the graft is discharged from the tubular tip, the beveled edge 98 of the tubular tip 96 promotes the graft 74 A progressive solution. The edge 98 of the tubular tip 96 is preferably beveled to 45 °. As indicated generally at 100. The narrow tubular tip 96 of the instrument 78 runs from the edge 98 of the tubular tip to the funnel. The narrowest end of 94 is preferably curved along its longitudinal axis. The curvature of the tubular tip 100 facilitates the operation of the instrument 78 in the eye, especially the instrument to a curved wall between the eyes, between the retina 32 And the supporting tissue 84. The radius of curvature 100 of the tubular tip will depend on the method of operation and the radius of curvature of the host eye. The instrument 78 further includes a plunger member 85 to help the graft pass through The narrow tubular tip 96. As shown in FIG. 18, the plunger member 85 is preferably a thin tubular post that is received in the open end 92 of the tubular body 90. The paper size is suitable for China National Standard (CNS) A4 Specification (210X297 Mm) 1 _ Order I (Please read the notes on the back before reading and writing this page) Printed by the Employees 'Cooperatives of the Central Standards Bureau of the Ministry of Economy 524681 Printed by the Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economy Α7 Β7 V. Description of the invention ( 17) Plug 86, so the relative advancement of the plunger about the tubular body passing through the funnel 94 and entering the tubular tip 96 drives the coiled fine, cellular structure 76 through the funnel of the tubular body and through the funnel The tubular tip of the instrument 78. In order to reduce the damage to the fragile cell structure 76 caused by the direct connection between the plunger 86 and the cell structure, the coiled cell structure 76 is Before being inserted, the spacer 90 is inserted into the open end 92 of the reputation body 90 and made of gelatin foam 102 or other soft and compressible material to avoid direct contact with the plunger 86. The gelatin foam 102 is guided on top of the coiled cell structure 76 and thus protects the coil from direct contact with the mechanical plug. Gelatin foam is satisfactory because it is semi-solid and non-toxic. The plunger 86 projects a sufficient distance from the open end 92 of the tubular body 90 so that the protruding end 88 of the plunger can be manipulated, even if the tubular tip 96 of the instrument 78 is false in the eye. The preferred method of operating the instrument 78 is once the tubular tip 96 with the coiled cell structure 76 inside is properly placed within the subretinal area 80 of the eye, and the plunger 86 is operated to remove the instrument from the instrument. The tubular tip 96 discharges the coiled cell structure 76. Although the plunger 86 provides maximum control of the discharge of the scroll 76 into the eye, due to the increased possibility of damage to the scroll 76, some care must be taken when operating the plunger. Alternatively, the plunger member 85 may include a member (not shown) for applying fluid pressure to the contents of the tubular body 90. In this regard, the open end 92 of the tubular body is connected to a fluid source under pressure. Fluid can be selectively supplied via a line connected to the open end 92 of the instrument 78 to replace its contents. The fluid can be viscous (such as 2¾ of carboxymethyl cellulose 21 — this paper size applies to China National Standard (CNS) A4 specifications (210X297 mm) (please read the precautions on the back before writing the page on this page)) · 524681 A7 B7 printed by the Consumer Cooperatives of the China Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (18) Vitamin) or non-sticky. With regard to the latter in particular, it may be desirable to place gelatin foam or some relatively soft spacer material within the tube to function as a mechanical plunger to separate the fluid from the cell structures implanted by the positive wave. As previously discussed, gelatin foam is satisfactory because it is semi-solid and will dissolve harmlessly if it is discharged from the instrument. Although the use of fluid pressure as the plunger member 85 significantly reduces the possibility of damage to the scroll 76, it also results in a significant reduction in the degree of control of the discharge of the scroll 76 from the instrument 78. As shown and described in patent application serial number 07 / 566,996, which is hereby incorporated by reference as a reference, the instrument can incorporate a number of features to facilitate a special surgery. As shown in Figure 17, the instrument may include a lumen 108 parallel to the instrument 78. As used herein, lumen 108 means any tubular container, whether it is provided separately or formed as a channel on the outside of the instrument 78. The lumen 108 has a distal tip 110 generally adjoining the tubular tip 96 of the instrument 78 and preferably slightly advanced relative to the tubular tip. The base end 112 of the lumen 108 is far from the distal end 110 and may be provided with a connector for fluid connection under pressure. Thus, the lumen 108 may discharge a fluid stream from its distal end 110 to create a fluid space before the instrument 78. The tubular tip 96 of the instrument 78 usually follows the passage opened by the fluid, thereby minimizing direct contact between the instrument and ocular tissue. The distal end 110 of the lumen 108 may be beveled 45 ° to facilitate the advancement of the instrument 78, even when the fluid is not being discharged from the lumen. The trailing end 110 is preferably chamfered to 45 °. The fluid discharged from the lumen 108 can be-22-This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) gutter (please read the precautions on the back before writing this page) Printed by the Consumer Standards Cooperative of the Central Bureau of Standards 524681 A7 B7 V. Description of the invention (19) A physiological saline solution or some other fluid that will not damage the eye tissues. Different examples such as antioxidants, anti-inflammatory agents, anti-mitotic agents, and local anesthetics can be provided in the fluid for treatment of the eye or implanted tissue. Depending on the type of surgery, the instrument may also include a fiber optic fiber (not shown) that extends substantially parallel to the lumen 108 and is placed between the lumen and the tubular body 90. The fiber optic fiber facilitates the operation of the instrument 78 and the proper placement of the graft 74 in two ways: a light source can be provided at the base of the optical fiber around which the fiber is provided at the tubular tip 96 of the instrument 78 Light to aid in visual inspection procedures through the pupil; optionally, a lens may be provided at the base end of the fiber optic fiber so that the fiber can be used for direct observation at the tubular tip of the instrument. In addition, the fiber can allow laser light to burn to control subretinal bleeding. The instrument 78 may further include a second lumen (not shown) that extends generally parallel to the first lumen 108 and is placed between the lumen 103 and the tube 90. The second lumen allows aspiration of material originating from the tubular tip 96 of the instrument 78. The base end of the lumen can be connected to a suction source to remove excess fluid and debris. The instrument 78 may further include a pair of wires (not shown. (Out), which terminates in the electrode at the end. The electrode allows cauterization of the blood vessels. The base end of the lead can be connected to a power source to seal a ruptured blood vessel. The wire can also be incorporated into the wall of the tubular body 90 of the instrument 78. Of course, two or more of the features described in the specific examples of these options may be combined together if necessary in the case to be determined. The method of transplanting a volute 76 to the subretinal area of one eye includes a 2 3-this paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm). Thread (please read the precautions on the back before dreaming) (This page) Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 524681 A7 _ B7___ V. Description of the invention (20) A transplantable substance (such as retinal pigment epithelium, choroid, Bruch's membrane and / or Retinal cells) combined into a graft 74. It will be understood that the implantable substance can be formed into a graft without a gelatin carrier sheet and still fall within the scope of the present invention. Preferably, however, the graft is combined with a carrier sheet 70. The transplantation method provides placement of the transplant 74 into the instrument at the open end 92 of the tubular body 90, and the graft 74 engages the inner wall of the tubular body. One end 73 of the graft 74 is first placed in the open end 92 of the tubular body 90, so the carrier 70 will be coiled so that the outer segment surface of the photoreceptor layer 54 faces the vortex formed. The outer side of the winding curve 77 of the object 76, so the vortex will be discontinued in the subretinal region so that the outer segment surface of the photoreceptor layer faces the pigment epithelium layer 84 of the host eye. The tubular tip 96 of the instrument 73 is capped 118 and the tubular body 90 is filled with a viscoelastic fluid 120 which facilitates the advancement of the graft into the conical channel or funnel. The graft slides into the funnel to engage a gradually narrowing tapered surface, which causes the graft to coil gradually. When the inner wall of the funnel 94 is narrow enough, the inverted edge 72 of the carrier sheet 70 becomes tight, and one side 72 of the sheet 70 slides below the other side of the carrier sheet. This is partly due to the Caused by cut sides. When the side edges become tight, the cut edges 72 prevent any wrinkling of the carrier sheet 70. In an optional specific example shown in FIG. 21, when the inner wall of the instrument 78 is narrow enough to bring the sides 72 of the graft 74 close to each other, the slope 99 guides one bit on the other side The lower side 72 starts the coiling of the scroll 76. At a certain point in the funnel 94, the winding 77 of the coiled material 76 was sufficiently compressed so that the elastic fluid 120 could no longer pass through the funnel one 24 — the paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) I —— ^ ------ see ------ 1T ------- ^ (Please read the notes on the back before filling this page) ♦ Central Standards Bureau of the Ministry of Economic Affairs Printed by Employee Consumer Cooperative 524681 A7 B7 V. Description of Invention (21) Force the coil. A gelatin foam spacer 102 is placed on the top of the coil 76, a spherical body 104 is placed on the instrument. A vacuum is created on the top end of the open end 92 of the device, and the fluid 120 and the vortex remain in the device 78. An injection needle 106 can be inserted through the spheroid 104 to inject more fluid 120 as needed. The plunger 86 is inserted into the open end 92 of the tubular body 90 via the spherical body 104 and is operated to contact the gelatin foam spacer 102. The plunger 86 is carefully advanced to force the graft 4 I, 74 through the funnel 94 to further coil the graft into a volute 76 and enter and pass through the tubular tip 96. Prepare the eyes of the host to reduce bleeding and surgical contusion. The scleral ciliary body approach to the subretinal space (Figure 20) is preferred, but other approaches can also be used, such as those crossing the cornea and sclera. Make a small incision in the ciliary body, and insert a surgical instrument 78 (about 0. 75min-about 2. 0mm). After the vitrectomy, a cooled and balanced saline solution is injected into the vitreous chamber of the eye 112 through a second ciliary body entrance to cool the eye to prevent the carrier sheet 70 of the vortex 76 from being surgically removed. Dissociation during processing. By making a mouth in the retina and injecting a balanced saline solution into the subretinal area to form a vesicle 80 at the transplant site of the retina 82, the retina 82, which is part of the transplant site, is lined from pigment epithelial cells δ4 lifts off. If the instrument 78 includes a lumen 108, the retina is subjected to the thermodynamic force of a perfusate (such as a physiological saline fluid, carboxymethyl cellulose, or 1-2¾ glacial acid) discharged from the lumen. The pupae were peeled to form a small bubble 80. Advantageously, the fluid may additionally contain antioxidants, anti-inflammatory agents, anesthetics, or it will reduce the host's retina 82. The paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) — order (please first Read the notes on the back and continue on this page.) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 524681 A7 B7 V. Description of the invention (22) Reagents required. An instrument 73 with the scroll 76 at its tubular tip 96 is inserted into the subretinal space through the ciliary body entrance, the vitreous chamber. As illustrated in Figure 20, the operation of the instrument 78 in turn causes the edge of the tubular tip 96 to be in line with the incision 122 of the vesicle 80. The entire tip of the instrument 78 is inserted into the vesicle 80, and the scroll 76 is discharged by carefully pushing the plunger. The scroll 76 is discharged from the chamfered edge 98 of the tubular tip 96 and recorded in its inherent solution, and when it is ejected from the chamfered edge, the coil is unwound, so the light The outer segment of the receptor layer 54 faces the pigment epithelium layer 84. If the volute 76 is not completely disentangled, the implant 74 may be fully disentangled using a microtooth pick. The vesicles 80 are then evacuated by fluid in the vesicles or evacuated by tenipa-nade, so that the graft 74 is placed in a sandwich arrangement near the retina 82 and pigmented epithelial lining 84 as desired. Location. The incision 122 of the vesicle 80 can be closed by burning. The gelatin carrier sheet 70 will dissolve once near normal body temperature. The edges of this scleral incision were removed after tweezers were removed and sutured using standard ophthalmic methods. As shown and described in the parent application serial number 07 / 566,996, a transchoroid, sclera, and corneal surgery method can be used as an alternative embodiment of the ciliary body margin method described above. Except for the point of entry, the surgical technique is substantially the same as the method described above. Based on the above, it can be seen that several objects of the present invention are achieved and other benefits are obtained. Due to the various changes in the above-mentioned surgical instruments, material composition and methods,-26-This paper size applies to Chinese National Standard (CNS) A4 (210x297 mm) f (Please read the precautions on the back and write this page first) -Binding · Order 524681 A7 B7 V. Explanation of the invention (23) Without departing from the scope of the present invention, all matters included in the above description or shown in the accompanying drawings will be covered. It is considered to be a phantom to illustrate the invention and not to limit the invention. (Please read the precautions on the back before writing this page.) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 7 2 This paper size applies to China National Standard (CNS) A4 (210X 297 mm)