WO2010137879A2 - Cellule souche de plante dérivée d'un cambium de la famille des salicaceae et procédé permettant de l'isoler et de la cultiver - Google Patents

Cellule souche de plante dérivée d'un cambium de la famille des salicaceae et procédé permettant de l'isoler et de la cultiver Download PDF

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WO2010137879A2
WO2010137879A2 PCT/KR2010/003342 KR2010003342W WO2010137879A2 WO 2010137879 A2 WO2010137879 A2 WO 2010137879A2 KR 2010003342 W KR2010003342 W KR 2010003342W WO 2010137879 A2 WO2010137879 A2 WO 2010137879A2
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cambium
stem cells
willow
derived
culture
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Korean (ko)
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WO2010137879A3 (fr
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홍남주
진영우
이은경
홍선미
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Unhwa Corp
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Unhwa Corp
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/76—Salicaceae (Willow family), e.g. poplar
    • 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
    • C12N5/04—Plant cells or tissues
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts

Definitions

  • the present invention relates to stem cells derived from the cambium of the family Salicaceae and to a method of isolation culture.
  • plant-derived useful substances have physiological activities such as antiviral, antibacterial, anticancer and antioxidant ability, they are attracting attention as an ideal resource for development as a new drug, and the relationship between chemical structure and activity of many plant-derived substances Researches are actively underway to identify the problem.
  • bioactive substances are difficult to develop as pharmaceuticals, and the main reasons thereof are as follows.
  • dedifferentiation process is essentially preceded to convert them into cell lines having dividing ability.
  • the dedifferentiation process means dismantling a state in which a tissue or cell has already been differentiated to perform a specific function when cultured using a tissue or organ of a plant.
  • severe variations in cell lines can occur due to chromosomal variations.
  • the production of useful substances through plant cell culture can be industrialized only if the rapid cell proliferation and high metabolite production capacity are maintained stably for a long period of culture, but most cell lines undergo numerous mutations different from the original by passage culture. You will come across. Therefore, in order to overcome such a mutation problem, a method for obtaining a genetically stable cell line in the production of useful substances through plant cell culture was urgently needed.
  • Some of the present inventors have developed a method of inducing callus using only the layer formed from the stem of the plant (Korean Patent No. 0533120), but this patent simply uses a stem forming layer of a tree plant to callus. I just induced.
  • Callus (Callus) is a tissue formed by dedifferentiation, so the patent has a problem that withholds the problem of variation by dedifferentiation.
  • some of the inventors of the present invention have developed the invention of International Patent Application No. PCT / KR2006 / 001544 as a method of providing a genetically stable cell line capable of reliably proliferating and solving the problem of mutation by dedifferentiation.
  • the poplar tree belonging to the willow family is widely known as a useful plant, and from this, it was required to solve the problem of mutation by dedifferentiation and to obtain a cell line with high genetic stability capable of stably proliferating.
  • the present inventors have made efforts to provide useful plant cell lines by isolating stem cells derived from the cambium of the cambium, and as a result, the stem cells are derived from the cambium of the willow, and the stem cells stably proliferate, It confirmed that there was no, and completed this invention.
  • the present invention provides a stem cell derived from the cambium of Willow family derived from the cambium of the family Salicaceae , characterized in that the natural undifferentiated cells have not undergone dedifferentiation.
  • the present invention also provides a method for separating stem-derived stem cells of the family Salicaceae comprising the following steps:
  • the present invention also provides an anti-inflammatory composition containing any one or more of the stem cells derived from the cambium of the willow, its extract, its lysate and its culture.
  • the present invention also provides a functional food for preventing or improving inflammation containing any one or more of the stem cells derived from the cambium of the willow, its extract, its lysate and its culture.
  • 1 is a photograph of the formation layer in the cross section of the material plant (poplar tree).
  • Figure 2 is a photograph of the induction and separation of stem cells according to the present invention
  • 2A is a photo of the stem cell induction medium containing the cambium layer
  • 2B shows the separation of the cambium-derived stem cells and heterogeneous other callus Photograph
  • 2C is a state in which only stem cell-derived stem cells were isolated and cultured for 3 weeks.
  • Figure 3 is a photograph of a solid culture of callus induced in the bark tissue of stem cells and poplar trees according to the present invention.
  • Figure 4 is a micrograph observing the degree of cell aggregation of the callus (B) induced in stem cells (A) and poplar bark tissue of the present culture process (x100).
  • FIG. 5 is a micrograph of a stem cell (A) and a callus (B) derived from a poplar bark tissue according to the present invention, and the scale bar at the bottom left is 25 ⁇ m.
  • Figure 6 is a micrograph of the stem cells (A) and the callus (B) derived from the poplar bark tissue according to the present invention observed after staining the vacuole using Neutral red, the scale bar at the bottom right is 25 ⁇ m .
  • Figure 7 is a photograph of the mitochondria of stem cells (A) and callus (B) induced in poplar bark tissues according to the present invention.
  • FIG. 8 is a graph showing the growth rate ratio of the callus (callus) induced in stem cells and poplar bark tissues according to the present invention.
  • FIG. 9 is a photograph showing the results of 14 days of culture in a 3L air-lift bioreactor (A) and a 20L air-lift bioreactor (B).
  • FIG. 10 is a graph showing the cell survival rate after cryopreservation of callus induced in stem cells and poplar bark tissues according to the present invention.
  • FIG. 11 is a graph showing the degree of NO generation inhibition according to the concentration of the stem cell extract according to the present invention when performing the NO analysis.
  • FIG. 12 is a gel photograph showing the degree of inhibition of COX-2 gene expression according to the concentration of the stem cell extract according to the present invention (A: 0.5g / ml, B: 1mg / ml, C: 2mg / ml).
  • the vascular "forming layer” is a lateral meristem located in the vascular tissue of the plant and is located in the stem and root.
  • the activity of the cambium causes hypertrophy of the plant, resulting in the presence of large vegetation with more than 11,000 years of age.
  • the vascular forming layer originates from the procambium, so that it can be conveniently divided into gradually divided mitotic tissues while maintaining mitotic continuity (Phytomorphology, co-author of Lee Jae-du et al., Academy Book, Chapter 10, 1993), Bonn
  • the formation layer is interpreted to include a formation layer. It is obvious that such a formation layer and a typical formation layer are the same primary fission structure, and the same effect will be acquired by using a formation layer and a typical formation structure in this invention.
  • Plant “stem cells” as used herein refer to innate undifferentiated cells that are genetically more stable without undergoing dedifferentiation.
  • cracked material refers to a cell lysate obtained by crushing a cell by a chemical method or a physical method using a detergent or the like
  • extract of a cell line is a material obtained by dissolving the cells in a solvent and using distillation or evaporation. Can be concentrated.
  • the cell line "culture medium” means a cell culture solution remaining after culturing the cells, excluding the cells.
  • culture is a substance containing a culture medium and / or a cultured cell line, wherein the cultured cell line is a concept including all cell lines that are differentiated by culture conditions or have improved production and / or secretion capacity of useful materials. .
  • innately undifferentiated refers to maintaining a pre-differentiation state, rather than being present in an undifferentiated state through a dedifferentiation process.
  • callus refers to cells or cell masses (PNAS, 99 (25): 15843, 2002) that have not been differentiated through dedifferentiation.
  • the present invention relates to stem cells derived from the cambium of Willowaceae in one aspect, which are innate undifferentiated cells derived from the cambium of Willowaceae and have not undergone dedifferentiation.
  • having a large number of vacuoles means having a plurality of vacuoles two times or more as compared to dedifferentiated callus and the like.
  • the stem cells according to the present invention has a small vacuole in size compared with the callus of the willow family.
  • having "a large number of advanced forms of mitochondria” refers to having more than twice the number of mitochondria that actively move under a microscope compared to the dedifferentiated callus of the willow family.
  • the stem cells may be characterized by having at least two or more of the properties of (a) to (e), preferably at least three of the properties of (a) to (e) It may be characterized by having at least four characteristics, more preferably at least four of the characteristics of the above (a) to (e). In addition, in the present invention, the stem cells may be characterized by having all the properties of (a) to (e).
  • the induction of differentiation of stem cells derived from the cambium cambium has a differentiation ability to differentiate into tracheary elements. Characterizing the plant stem cells has a differentiation ability (pluripotency) in addition to the self renewal ability (self renewal), whereby it was confirmed that the willow-derived cell derived from the stem cells according to the present invention.
  • stem cells according to the present invention comprises the steps of (a) obtaining a cambium-containing tissue from the willow plant; (b) culturing the obtained cambium-containing tissue in a medium; And (c) obtaining the cambium-derived stem cells by separating the cells from the cambium, wherein step (b) is a step of culturing the cambium-containing tissue to grow from the cambium. It may be characterized by inducing the formed layer, the step (c) may be characterized in that to obtain a cambium-derived stem cells by separating the cultured cambium.
  • the step (b) may be characterized by culturing in a medium containing auxin (auxin), wherein, as the auxin is used NAA ( ⁇ -Naphtalene acetic acid) or IAA (Indole-3-acetic acid)
  • the auxin may be characterized in that it is included at a concentration of 1 ⁇ 5mg / l.
  • the step (c) may be characterized in that to obtain a cambium-derived stem cells by separating the cultured cambium layer from the callus layer that is amorphously proliferated from portions other than the cambium.
  • the cambium-derived stem cells of the poplar are isolated from the poplars of the willow and aspen, and the present invention is preferably characterized in that the cambium-derived stem cells of the genus Aspen is more preferred. It may be characterized that the cambium-derived stem cells.
  • the anti-inflammatory effect of the cambium-derived stem cells from the poplar which is obtained from the above-mentioned Willowaceae aspen, was confirmed, and the present invention, in another aspect, the stem cells derived from the cambium of the willow family, extracts thereof.
  • the present invention relates to an anti-inflammatory composition containing any one or more of its lysate and its culture.
  • the culture in the present invention the stem cells as an eliminator 3 to 5% by weight of raw sugar or sugar; Or further culturing in a medium containing any one or more of methyl jasmonate, chitosan, phenylalanine, benzoic acid, ABA, salicylic acid, and sodium acetate. It can be characterized in that obtained by performing.
  • the medium is 3 to 5% by weight of raw sugar or sugar; And methyl jasmonate, fungal extract, bacterial extract, yeast extract, chitosan, glucomanan, glucan, phenylalanine, benzoic acid, salicylic acid ), Arachonic acid, STS, mevalonalonate N-benzolyglycine, ABA, SNP, IPP, BHT, CCC, ethephon, hipuic acid, ammonium ceric nitrate, A substance selected from the group consisting of AgNO 3 , vanadyl sulfate, p-aminobenzoic acid, brassinosteroids, sodium alginate, sodium acetate It may be characterized in that the medium containing.
  • the present invention it is also possible to use a culture obtained by applying physical and chemical stress to the stem cells by treating ultraviolet rays, heat, ethylene, antifungal agents, antibiotics, heavy metal salts and high salt concentrations as an eliminator.
  • the extract may be extracted using a solvent selected from the group consisting of distilled water, alcohol such as ethanol, acetone, DMSO (dimethyl sulfoxide) and a mixed solvent thereof.
  • a solvent selected from the group consisting of distilled water, alcohol such as ethanol, acetone, DMSO (dimethyl sulfoxide) and a mixed solvent thereof.
  • the stem cell extract according to the present invention can be usefully used as a very excellent anti-inflammatory composition.
  • the composition containing the stem cells or the lysate exhibits an anti-inflammatory effect, as described above, it was confirmed that the stem cell extract has anti-inflammatory activity. It will be apparent to those skilled in the art that the stem cell itself, the lysate thereof, and the composition containing the culture according to the present invention can also exhibit anti-inflammatory activity and suppress inflammation.
  • An anti-inflammatory composition containing any one or more of stem cells, extracts thereof, lysates, and cultures thereof according to the present invention as an active ingredient, each of them alone or at least one pharmaceutically acceptable carrier, excipient or diluent It may be provided as a pharmaceutical composition, the stem cells and the like to be included in the pharmaceutical composition in an appropriate pharmaceutically effective amount according to the disease and its severity, the age, weight, health status, sex, route of administration and duration of treatment, etc. Can be.
  • pharmaceutically acceptable refers to a composition that is physiologically acceptable and that, when administered to a human, typically does not cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like.
  • carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • the pharmaceutical composition may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives.
  • Pharmaceutical compositions of the invention can also be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.
  • the formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders.
  • the present invention relates to a functional food for preventing or improving inflammation containing any one or more of stem cells, extracts thereof, lysates and cultures thereof according to the present invention as an active ingredient.
  • the term "functional food” means that the functionality of the food is improved by adding any one or more of the stem cells or the extract thereof, the lysate, and the culture according to the present invention to a general food.
  • the fragment comprising the formation layer prepared in Example 1-2 was cultured by denture on the stem cell induction medium (media 1) of Table 1.
  • Auxins such as NAA and IAA as growth regulators may be added to the medium at a concentration of 1 to 5 mg / L, preferably at a concentration of 2 mg / L. Cultivation was carried out in a dark room controlled at 25 ⁇ 1 °C.
  • FIG. 2C is a state in which only stem cell-derived stem cells were isolated and cultured for 3 weeks.
  • the bark sections of the poplar tree were sterilized for comparison and then cultured in the medium of Table 1, as a result, as shown in FIG. 3B, the bark sections form callus by dedifferentiation.
  • Callus derived from bark fragments was amorphous due to differences in the rate of division between cells, such as tetrahedral tissue, and showed unstable growth rate and easy browning.
  • Callus derived from browned and agglomerated bark was slowed down by the phenolic compounds it secreted and eventually became necrotic. That is, callus derived from bark from 6 months later was difficult to maintain and incubate.
  • FIG. 3A the cambium-derived stem cells of the poplar tree were stably maintained without change in cell growth rate, growth pattern and degree of aggregation during long-term culture, thereby enabling mass culture.
  • the stem cell-derived stem cells isolated in Example 1 were placed in a flask containing the liquid medium of the following ⁇ Table 2> and cultured in a rotary shaker of 100 rpm at 25 ⁇ 1 °C under dark conditions.
  • the subculture period was fixed at 2 weeks so that the cultured cells could always maintain high vitality in the algebraic state.
  • the stem cells according to the present invention when observed the degree of aggregation in the solid culture, is observed as shown in Figure 4A, a large number of single cells during the solid culture process Included, some were confirmed to exist as a small size cell aggregate. On the contrary, in the case of callus induced in the bark tissue of the control poplar tree, it was observed to aggregate as shown in FIG. 4B.
  • the stem cells according to the present invention when suspended in culture in medium 2 of Table 2, are present as a small cell aggregate having a size of 300 ⁇ m in the initial suspension culture, cells having a size of 450 ⁇ m or more in the initial suspension culture.
  • the cell aggregates were smaller than the callus showing the aggregates.
  • the callus was formed in a much more aggregated form, whereas the stem cells according to the present invention were confirmed to include a large number of single cells or a small cell aggregate.
  • stem cells according to the present invention was able to observe the morphological characteristics having a plurality of vacuole (vacuole).
  • This feature is a feature that appears due to causes such as pressure in the undifferentiated cells present in the plant, it was confirmed that the stem cells according to the present invention is in an undifferentiated state.
  • somatic cells callus derived from bark tissue
  • FIG. 5B such a feature could not be confirmed.
  • stem cells according to the present invention were able to identify a number of small vacuoles of red as shown in FIG. 6A, and the somatic cells of the poplar tree (bark) In the case of callus induced in the tissue, as shown in FIG. 6B, one large central vacuole was present.
  • stem cells according to the present invention was observed through the optical microscope BX41TF, as a result, it was able to observe a large number of mitochondria very active in terms of movement.
  • 7A shows that stem cells according to the present invention have a plurality of mitochondria, and arrows indicate mitochondria.
  • somatic cells callus derived from the bark tissue
  • FIG. 7B these characteristics could not be confirmed.
  • the cambium-derived stem cells according to the present invention have low sensitivity to shear stress in the bioreactor for mass culture, and thus, it was confirmed that rapid mass growth is possible in the bioreactor. Therefore, it was found that the cambium-derived stem cells according to the present invention have low sensitivity to shear stress as compared to the dedifferentiated callus of poplar.
  • cryopreservation was performed on the callus derived from the poplar bark tissue and the stem cells derived from the poplar forming layer according to the present invention.
  • Suspension culture is used for 6 to 8 days of culture, cryopreservative is a medium containing 0.5M glycerol (DUCHEFA, The Netherlands) and 0.5M DMSO (DUCHEFA, The Netherlands) and 1M sucrose (DUCHEFA, The Netherlands) , 5 ml cryovial (Duran, USA).
  • the amount of cell inoculation treated with cryopreservative is 200 mg / ml.
  • the cryopreservative treated suspension cells were kept in a freezer for 30 minutes and then stored in a deep freezer for 3 hours and then frozen by immersion in liquid nitrogen.
  • the cultured cells kept in liquid nitrogen for 20 minutes or more were taken out for thawing and thawed in a 40 ° C. constant temperature water bath for 1 to 2 minutes.
  • the cell suspension was used aseptic funnel and filter paper. The filtered cells were applied on solid growth media containing filter paper, stabilized at room temperature for 30 minutes, and then transferred back to fresh solid growth media.
  • the characteristics that characterize the plant stem cells have a pluripotency in addition to self renewal (self renewal).
  • self renewal self renewal
  • the conduit element differentiation of the stem cells derived from the poplar tree it was confirmed that the conduit element was differentiated from the cambium stem cells as a result of culturing in the condition of 25 ⁇ 1 °C, dark conditions under the growth medium added MS.
  • Stem cells suspended in culture for 14 days were collected as in Example 2, and the experiment was performed by dividing into two treatment groups. That is, (1) the cell phase suspended in culture for 14 days (growth phase), (2) 3 to 5% by weight (g / L) of methyl jasmonate and 100 ⁇ M of methyl jasmonate were added to sterile water in the cell line suspended in culture for 14 days Cancer cells were used for 14 days in the elicitation phase (elicitation phase).
  • Each of the two cell lines was removed from the culture solution and then lyophilized in 2 g of lyophilized lyophilized cell line (Dry) with 50 ml of 80% ethanol at 15 ° C. for 48 hours. After the lysis, the extract powder obtained by lyophilization of the supernatant by centrifugation at 3,000 rpm for 20 minutes was dissolved in PBS, and ethanol extracts of the two cell lines were obtained and used, respectively.
  • the NO assay is a method of measuring macrophage activity and is an indirect activity measurement method associated with phagocytosis, which is one of the important functions of macrophages.
  • the ethanol extract of the stem cells derived from the poplar forming layer of Example 3 was measured by the amount of NO dissolved in the supernatant to inhibit the NO production generated when LPS (Lipopolysaccharide) was treated in RAW264.7 cells.
  • the ethanol extract of the poplar forming layer-derived stem cells were found to inhibit NO production induced by 22% at 1mg / ml, 42% LPS at 1.3mg / ml, 5mg / ml In concentration, NO production induced by LPS was confirmed to be inhibited by 80%.
  • RAW264.7 cells (2X10 6 / ml) and samples were collected in 6 well culture plates (A: 0.5g / ml, B: 1mg). / ml, C: 2mg / ml) for 30 minutes pre-culture, cells treated with LPS (1ug / ml) for 24 hours to collect the total RNA was isolated from TRIzol to synthesize cDNA. Then, PCR was performed using COX-2 Primer.
  • mouse COX2 F 5’-AAGAAGAAAGTTCATTCCTGATCCC-3 ’(SEQ ID NO: 1)
  • mouse COX2 R 5'-TGACTGTGGGAGGATACATCTCTCC-3 '(SEQ ID NO: 2)
  • 100 mg of the stem cell extract prepared in Example 3 was mixed according to the conventional tablet preparation method by mixing corn starch 100 mg, lactose 100 mg and magnesium stearate 2 mg.
  • 500 mg of the stem cell extract prepared in Example 3 was filled into a soft gelatin capsule to prepare a capsule.
  • 100 ml of syrup was prepared according to a conventional method of preparing a liquid formulation with the content of 1 g of stem cells obtained in Example 1, 10 g of isomerized sugar, 5 g of mannitol, and an appropriate amount of purified water.
  • Example 2 After dissolving 200 mg of the stem cells obtained in Example 1 in 96 ml of water, 500 mg of vitamin C as an adjuvant, 1 g of citric acid and oligosaccharides as coppers were added, and 0.05 g of sodium benzoate as a preservative, followed by purified water. The amount was added to 100ml to prepare a functional beverage.
  • the stem cells derived from the cambium of the Willow family according to the present invention are separated into an undifferentiated state without dedifferentiation process, and thus, even during long-term culture, the stem cells are stably maintained without variation in cell growth rate and growth pattern and thus useful for mass culture.
  • the cambium-derived stem cells of the present invention according to the present invention has an effect of inhibiting the generation of NO generated when LPS (Lipopolysaccharide) treatment and the effect of inhibiting the gene expression of COX-2 inflammatory cytokines, excellent anti- It can be usefully used as an inflammatory composition.

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Abstract

La présente invention concerne une cellule souche dérivée d'un cambium de la famille des Salicaceae et un procédé permettant de l'isoler et la cultiver. La cellule souche dérivée d'un cambium de la famille des Salicaceae selon la présente invention est avantageuse étant donnée qu'elle est isolée dans un état non différencié sans passer par un processus de dédifférenciation et maintenue de manière stable sans provoquer de variation de la vitesse de croissance cellulaire et du type de croissance même pendant une culture de longue durée, ce qui permet ainsi de réaliser une culture de masse. De plus, la cellule souche dérivée d'un cambium de la famille des Salicaceae selon la présente invention présente les effets de supprimer la génération de NO pendant le traitement par Lipopolysaccharide (LPS) et de supprimer l'expression des gènes de la cytokine pro-inflammatoire COX-2 et peut ainsi être utilisée avec intérêt en tant qu'excellente composition anti-inflammatoire.
PCT/KR2010/003342 2009-05-26 2010-05-26 Cellule souche de plante dérivée d'un cambium de la famille des salicaceae et procédé permettant de l'isoler et de la cultiver Ceased WO2010137879A2 (fr)

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KR10-2009-0045955 2009-05-26

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WO2010137879A2 true WO2010137879A2 (fr) 2010-12-02
WO2010137879A3 WO2010137879A3 (fr) 2011-04-14

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CN116589606A (zh) * 2022-12-01 2023-08-15 中国药科大学 一种丁酰化酵母葡聚糖及其制备方法、应用

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CN115968731A (zh) * 2023-02-09 2023-04-18 甘肃农业大学 一种用外源一氧化氮和水杨酸缓解玉米幼苗盐胁迫的方法

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013102882A2 (fr) 2012-01-05 2013-07-11 L'oreal Utilisation cosmétique de cellules végétales dédifférenciées
US10463603B2 (en) 2012-01-05 2019-11-05 L'oreal Cosmetic use of dedifferentiated plant cells
CN116589606A (zh) * 2022-12-01 2023-08-15 中国药科大学 一种丁酰化酵母葡聚糖及其制备方法、应用
CN116589606B (zh) * 2022-12-01 2024-05-28 中国药科大学 一种丁酰化酵母葡聚糖及其制备方法、应用

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