EP0230430A4 - CULTURE MEDIUM FOR FERTILIZATION -i (IN VITRO) AND TRANSFER OF EMBRYOS. - Google Patents

CULTURE MEDIUM FOR FERTILIZATION -i (IN VITRO) AND TRANSFER OF EMBRYOS.

Info

Publication number: EP0230430A4
Authority: EP; European Patent Office
Prior art keywords: sodium; potassium; culture medium; ions; chloride
Prior art date: 1985-06-12
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.): Withdrawn

Application number

EP19860903165

Other languages

German (de)

English (en)

French (fr)

Other versions

EP0230430A1 (en

Inventor

Patrick James Qulnn

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Luminis Pty Ltd

Original Assignee

Luminis Pty Ltd

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

1985-06-12

Filing date

1986-06-12

Publication date

1989-03-09

1986-06-12 Application filed by Luminis Pty Ltd filed Critical Luminis Pty Ltd

1987-08-05 Publication of EP0230430A1 publication Critical patent/EP0230430A1/en

1989-03-09 Publication of EP0230430A4 publication Critical patent/EP0230430A4/en

Status Withdrawn legal-status Critical Current

Links

239000001963 growth medium Substances 0.000 title claims abstract description 32
230000004720 fertilization Effects 0.000 title claims abstract description 18
238000000338 in vitro Methods 0.000 title claims abstract description 16
210000001161 mammalian embryo Anatomy 0.000 title description 7
UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims abstract description 38
DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims abstract description 20
229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 19
GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 19
229910000030 sodium bicarbonate Inorganic materials 0.000 claims abstract description 19
229910001415 sodium ion Inorganic materials 0.000 claims abstract description 19
FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 18
229910001414 potassium ion Inorganic materials 0.000 claims abstract description 18
210000000287 oocyte Anatomy 0.000 claims abstract description 12
CYDQOEWLBCCFJZ-UHFFFAOYSA-N 4-(4-fluorophenyl)oxane-4-carboxylic acid Chemical compound C=1C=C(F)C=CC=1C1(C(=O)O)CCOCC1 CYDQOEWLBCCFJZ-UHFFFAOYSA-N 0.000 claims abstract description 10
WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 10
239000007836 KH2PO4 Substances 0.000 claims abstract description 10
BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 claims abstract description 10
LLSDKQJKOVVTOJ-UHFFFAOYSA-L calcium chloride dihydrate Chemical compound O.O.[Cl-].[Cl-].[Ca+2] LLSDKQJKOVVTOJ-UHFFFAOYSA-L 0.000 claims abstract description 10
239000008103 glucose Substances 0.000 claims abstract description 10
235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 10
229960003531 phenolsulfonphthalein Drugs 0.000 claims abstract description 10
239000001103 potassium chloride Substances 0.000 claims abstract description 10
235000017557 sodium bicarbonate Nutrition 0.000 claims abstract description 10
239000001540 sodium lactate Substances 0.000 claims abstract description 10
235000011088 sodium lactate Nutrition 0.000 claims abstract description 10
229940005581 sodium lactate Drugs 0.000 claims abstract description 10
229940054269 sodium pyruvate Drugs 0.000 claims abstract description 10
239000011780 sodium chloride Substances 0.000 claims abstract description 9
229930182555 Penicillin Natural products 0.000 claims abstract description 5
JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims abstract description 5
229940049954 penicillin Drugs 0.000 claims abstract description 5
CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 28
WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 27
229910052943 magnesium sulfate Inorganic materials 0.000 claims description 18
239000000203 mixture Substances 0.000 claims description 13
238000000034 method Methods 0.000 claims description 10
235000019341 magnesium sulphate Nutrition 0.000 claims description 9
235000011164 potassium chloride Nutrition 0.000 claims description 9
239000001110 calcium chloride Substances 0.000 claims description 8
229910001628 calcium chloride Inorganic materials 0.000 claims description 8
XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 8
KZDCMKVLEYCGQX-UDPGNSCCSA-N 2-(diethylamino)ethyl 4-aminobenzoate;(2s,5r,6r)-3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid;hydrate Chemical group O.CCN(CC)CCOC(=O)C1=CC=C(N)C=C1.N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 KZDCMKVLEYCGQX-UDPGNSCCSA-N 0.000 claims description 5
DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 5
FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical compound [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 claims description 5
239000011734 sodium Substances 0.000 claims description 5
NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 claims description 4
150000001875 compounds Chemical class 0.000 claims description 3
WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 abstract 1
239000012530 fluid Substances 0.000 description 20
239000002609 medium Substances 0.000 description 19
230000035935 pregnancy Effects 0.000 description 9
230000000977 initiatory effect Effects 0.000 description 6
IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 4
CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 4
230000013020 embryo development Effects 0.000 description 4
239000011591 potassium Substances 0.000 description 4
229910052700 potassium Inorganic materials 0.000 description 4
229910052708 sodium Inorganic materials 0.000 description 4
210000002257 embryonic structure Anatomy 0.000 description 3
QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
210000002459 blastocyst Anatomy 0.000 description 2
239000001569 carbon dioxide Substances 0.000 description 2
229910002092 carbon dioxide Inorganic materials 0.000 description 2
239000007789 gas Substances 0.000 description 2
239000011521 glass Substances 0.000 description 2
239000012528 membrane Substances 0.000 description 2
229910052757 nitrogen Inorganic materials 0.000 description 2
239000001301 oxygen Substances 0.000 description 2
229910052760 oxygen Inorganic materials 0.000 description 2
235000004252 protein component Nutrition 0.000 description 2
238000003776 cleavage reaction Methods 0.000 description 1
238000011534 incubation Methods 0.000 description 1
BITYAPCSNKJESK-UHFFFAOYSA-N potassiosodium Chemical compound [Na].[K] BITYAPCSNKJESK-UHFFFAOYSA-N 0.000 description 1
230000010076 replication Effects 0.000 description 1
230000007017 scission Effects 0.000 description 1
230000035899 viability Effects 0.000 description 1

Classifications

- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0608—Germ cells
- C12N5/0609—Oocytes, oogonia
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/05—Inorganic components
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/34—Sugars

Definitions

the invention relates to culture media and more particularly to culture media for in vitro fertilization and embryo transfer.
n vitro fertilization of human oocytes, cleavage of embryos and embryo transfer require that a culture medium be used to support the embryo for a period of up to three or four days during the various processes necessary for fertilization and early incubation before embryo transfer and reimplantatlon.
the oocyte In the natural process of fertilization for a human oocyte the oocyte is supported within the mother within a fluid known as human tubal fluid andi t i s the object of the present invention to provide a culture medium as a synthetic human tubal fluid.
the present invention we have devised a synthetic culture medium which is believed to approximate human tubal fluid but with desirable additional components and variations in the actual composition, including ratios of concentrations of sodium ions to potassium ions.
the present invention may be said to reside in a culture medium for in vitro fertilization and embryo transfer comprising the following compounds in the following ranges of concentration;
the ratio of sodium ion concentration to potassium ion concentration is in the range from 28 to 32.
the ratio of concentrations of sodium ions to potassium ions is 29.3.
the synthetic human tubal fluid may have the following composition
the invention may be said to reside in a method of assisting with the in vitro fertilization of human oocytes including the step of handling the human oocytes in a culture medium, the culture medium being comprised of the compounds listed below in the range of compositions listed as follows;
the ratio of sodium ion concentration to potassium ion concentration is in the range of from 28 to 32.
the ratio concentration of sodium ions to potassium ions is 29.3.
the method includes the step of handling the oocytes in a culture medium comprising;
thei nvention may be said to reside in a culture medium for thei n vitro fertilization of human oocytesi ncluding sodium potassiumi ons wherein the ratio of sodium ions to potassium ions is i n the range of from 28 to 32.
the ratio of sodium ions to potassium ions is 29.3.
synthetic human tubal fluid culture medium is as given in Table 1 below (marked synthetic HTF).
the medium may be prepared by using rainwater which has been distilled in glass six times.
the bicarbonate-buffered medium is gassed for a minimum of five minutes with humidified 5% oxygen 5% carbon dioxide 90% nitrogen mixture and sterilized by passage through a 0.45-0.2 micrometre filter membrane (Millipore, Sydney, Australia or Amicon Sterilet Sydney, Australia) and then stored at 4°C for up to two weeks before use.
a minimum of six hours or preferably the day before being used the bicarbonate buffered medium is gassed again for two to three minutes with the same gas mixture as above and the protein component is added.
Tests have been carried out using both mouse embryo development in vitro and with initiation of human pregnancy in an endeavour to discover which components of the T6 medium and the synthetic human tubal fluid according to this invention might be responsible for observed differences in mouse embryo development in vitro and initiation of human pregnancies.
the results show that for human pregnancy initiation almost three times as many pregnancies occurred when fertilization and culture were carried out using the synthetic human tubal fluid of the present invention over the T6 medium.
the present invention therefore provides a synthetic human tubal fluid culture medium which is more than just a direct replication of naturally occurring human tubal fluid but has enhanced viability.
Potassium chloride (Kcl) 4.69 mM
the invention may be said to reside i n a culture medium for the in vitro fertilization of human oocytes i ncluding sodium potassium i ons wherein the ratio of sodium i ons to potassium ions i s in the range of from 28 to 32.
the ratio of sodium ions to potassiumi ons is 29.3.
synthetic human tubal fluid culture medium i s as given in Table 1 below (marked synthetic HTF).
the medium may be prepared by using rainwater which has been distilled in glass six times.
the bicarbonate-buffered medium is gassed for a minimum of five minutes with humidified 5% oxygen 5% carbon dioxide 90% nitrogen mixture and sterilized by passage through a 0.45-0.2 micrometre filter membrane (Millipore, Sydney, Australia or Amlcon Sterilet Sydney, Australia) and then stored at 4°C for up to two weeks before use.
a minimum of six hours or preferably the day before being used the bicarbonate buffered medium is gassed again for two to three minutes with the same gas mixture as above and the protein component is added.
Tests have been carried out using both mouse embryo development in vitro and with initiation of human pregnancy in an endeavour to discover which components of the T6 medium and the synthetic human tubal fluid according to this invention might be responsible for observed differences in mouse embryo development in vitro and initiation of human pregnancies.
the results show that for human pregnancy initiation almost three times as many pregnancies occurred when fertilization and culture were carried out

Landscapes

Life Sciences & Earth Sciences (AREA)
Health & Medical Sciences (AREA)
Engineering & Computer Science (AREA)
Biomedical Technology (AREA)
Genetics & Genomics (AREA)
Zoology (AREA)
Organic Chemistry (AREA)
Bioinformatics & Cheminformatics (AREA)
Biotechnology (AREA)
Chemical & Material Sciences (AREA)
Wood Science & Technology (AREA)
Developmental Biology & Embryology (AREA)
Microbiology (AREA)
Biochemistry (AREA)
General Engineering & Computer Science (AREA)
General Health & Medical Sciences (AREA)
Cell Biology (AREA)
Micro-Organisms Or Cultivation Processes Thereof (AREA)

EP19860903165 1985-06-12 1986-06-12 CULTURE MEDIUM FOR FERTILIZATION -i (IN VITRO) AND TRANSFER OF EMBRYOS. Withdrawn EP0230430A4 (en)

Applications Claiming Priority (2)

Application Number	Priority Date	Filing Date	Title
AU1009/85		1985-06-12
AUPH100985		1985-06-12

Publications (2)

Publication Number	Publication Date
EP0230430A1 EP0230430A1 (en)	1987-08-05
EP0230430A4 true EP0230430A4 (en)	1989-03-09

Family

ID=3771143

Family Applications (1)

Application Number	Title	Priority Date	Filing Date
EP19860903165 Withdrawn EP0230430A4 (en)	1985-06-12	1986-06-12	CULTURE MEDIUM FOR FERTILIZATION -i (IN VITRO) AND TRANSFER OF EMBRYOS.

Country Status (3)

Country	Link
EP (1)	EP0230430A4 (da)
DK (1)	DK66187A (da)
WO (1)	WO1986007377A1 (da)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number	Priority date	Publication date	Assignee	Title
JPH06197665A (ja) *	1993-01-07	1994-07-19	Norin Suisansyo Chikusan Shikenjo	体外受精用培地および体外受精方法
AU2707100A (en)	1998-11-30	2000-06-19	Ivf Sciences Colorado, Inc.	System and sequential culture media for in vitro fertilization
JP3856422B2 (ja) *	1999-01-05	2006-12-13	ニプロ株式会社	動物胚培養用培地
CN1922307A (zh)	2003-12-30	2007-02-28	财团法人首尔大学校产学协力财团	胚胎干细胞系及其制备方法
EP2787810A4 (en) *	2011-12-09	2015-05-27	Univ Missouri	Inorganic Pyrophosphates and Uses Thereof
CN111778202B (zh) *	2020-07-10	2022-10-28	深圳韦拓生物科技有限公司	一种颗粒细胞去除液及其制备方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number	Priority date	Publication date	Assignee	Title
US4473647A (en) *	1981-02-27	1984-09-25	Amf Inc.	Tissue culture medium
US4443432A (en) *	1981-10-05	1984-04-17	Alcon Laboratories, Inc.	Ophthmalic irrigating solution
JPS61502933A (ja) *	1984-06-22	1986-12-18	ビ−チ、リチャ−ド・エル	電解質溶液およびその生体外での使用

1986
- 1986-06-12 EP EP19860903165 patent/EP0230430A4/en not_active Withdrawn
- 1986-06-12 WO PCT/AU1986/000170 patent/WO1986007377A1/en not_active Ceased
1987
- 1987-02-10 DK DK066187A patent/DK66187A/da not_active Application Discontinuation

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BIOLOGICAL ABSTRACTS, vol. 81, 1986, no. 28226, Philadelphia, PA, US; R.K. NAZ et al.: "Successful human pregnancy following in vitro fertillization using frozen semen", & J. IN VITRO FERT EMBRYO TRANSFER 2(3), 143-145, 1985 *
BIOLOGICAL ABSTRACTS/RRM, no. 30096151; P. QUINN et al.: "Improved pregnancy rate in human in-vitro using a culture medium based on the composition of human tubal fluid", & 3RD ANNUAL SCIENTIFIC MEETING OF THE FERTILITY SOCIETY OF AUSTRALIA, BRISBANE, AUSTRALIA, Nov. 1984, CLIN REPROD FERTIL 1985 (RECD. 1986) vol. 3, no. 3, p220-221 *
CHEMICAL ABSTRACTS, vol. 81, no. 13, 30th September 1974, page 287, abstract no. 75696n, Columbus, Ohio, US; Y. TOYODA et al.: "Capacitation of epididymal spermatozoa in a medium with high potassium/sodium ratio and cyclic AMP for the fertilization of rat eggs in vitro", & J. REPROD. FERT. 1974, 36(1), 125-34 *
FERTILITY AND STERILITY, vol. 27, no. 6, 1976, pages 677-684; A. LOPATA et al.: "A method for collecting motile spermatozoa from human semen" *
IN VITRO, vol. 6, no. 2, 1970, pages 89-108; H.J. MORTON: "A survey of commercially available tissue culture media" *
See also references of WO8607377A1 *

Also Published As

Publication number	Publication date
DK66187D0 (da)	1987-02-10
DK66187A (da)	1987-03-26
WO1986007377A1 (en)	1986-12-18
EP0230430A1 (en)	1987-08-05

Legal Events

Date	Code	Title	Description
1987-06-20	PUAI	Public reference made under article 153(3) epc to a published international application that has entered the european phase	Free format text: ORIGINAL CODE: 0009012
1987-08-05	AK	Designated contracting states	Kind code of ref document: A1 Designated state(s): CH DE FR GB LI NL SE
1987-08-12	17P	Request for examination filed	Effective date: 19870617
1989-04-26	A4	Supplementary search report drawn up and despatched	Effective date: 19890309
1991-05-02	STAA	Information on the status of an ep patent application or granted ep patent	Free format text: STATUS: THE APPLICATION HAS BEEN WITHDRAWN
1991-05-15	18W	Application withdrawn	Withdrawal date: 19910331
1991-06-19	18D	Application deemed to be withdrawn	Effective date: 19910103
1991-06-19	R18W	Application withdrawn (corrected)	Effective date: 19910331
2007-08-08	RIN1	Information on inventor provided before grant (corrected)	Inventor name: QULNN, PATRICK, JAMES

Publication	Publication Date	Title
Boatman	1987	In vitro growth of non-human primate pre-and peri-implantation embryos
Whitten	1971	Nutrient requirements for the culture of preimplantation embryos in vitro
Wiley et al.	1986	Effect of potassium concentration, type of protein supplement, and embryo density on mouse preimplantation development in vitro
Stuart et al.	1969	Studies on the Growth in Culture of Plant Cells: IV. THE INITIATION OF DIVISION IN SUSPENSIONS OF STATIONARY-PHASE CELLS OF ACER PSEUDOPLATANUS L.
Lewontin	1970	The units of selection
Visvesvara	1980	Axenic growth of Giardia lamblia in Diamond's TPS-1 medium
Wright Jr et al.	1976	Successful culture in vitro of bovine embryos to the blastocyst stage
Maeda	1970	Influence of ionic conditions on cell differentiation and morphogenesis of the cellular slime molds
Keefer et al.	1982	Spontaneous activation of ovulated rat oocytes during in vitro culture
MAHI et al.	1973	The effects of temperature, osmolality and hydrogen ion concentration on the activation and acrosome reaction of golden hamster spermatozoa
JPS59232090A (ja)	1984-12-26	血清代替物を含む細胞培地
KR880700064A (ko)	1988-02-15	고무 폴리머라제 및 이의 제법과 용도
Brinster et al.	1979	Requirements for blastocyst development in vitro
EP0230430A4 (en)	1989-03-09	CULTURE MEDIUM FOR FERTILIZATION -i (IN VITRO) AND TRANSFER OF EMBRYOS.
Kaplan et al.	1978	Effect of elevated calcium concentration on fertilization of rat oocytes in vitro
Fildes et al.	1933	The nitrogen and vitamin requirements of B. typhosus
AU588339B2 (en)	1989-09-14	Culture media for in vitro fertilization and embryo transfer
Karn	1972	Sexual and asexual differentiation in Volvox obversus (Shaw) Printz, strains Wd3 and Wd7
Mahadevan et al.	1986	Growth of mouse embryos in bicarbonate media buffered by carbon dioxide, hepes, or phosphate
Spindle	1990	In vitro development of one-cell embryos from outbred mice: influence of culture medium composition
O'Kelley et al.	1961	Alkaline earth elements and zoospore release and development in Protosiphon botryoides
Maurizi et al.	1983	Morphological and functional characteristics of human temporal-bone cell cultures
WO2013087755A1 (en)	2013-06-20	Culture media for treating female gametes and embryos in medically assisted reproduction techniques
Yoshimatsu et al.	1988	Effects of Cations and Other Medium Components on the Zona‐induced Acrosome Reaction of Hamster Spermatozoa: (acrosome reaction/spermatozoa/cations/hamster)
Chang	1942	Studies on Entamoeba Histolytica: I. Effect of Hydrogen-Ion Concentration on Encystation of E. histolytica in Culture