Hadasit Medical Research Services & Development Limited
대리인 / 주소
Vorys, Sater, Seymour and Pease LLP
인용정보
피인용 횟수 :
7인용 특허 :
4
초록▼
The present application discloses methods expanding SCs in an undifferentiated state, the methods comprising incubating undifferentiated SCs in suspension within a culture system comprising basic medium and knockout serum replacement (KOSR). The methods may also be applicable for selective spontaneo
The present application discloses methods expanding SCs in an undifferentiated state, the methods comprising incubating undifferentiated SCs in suspension within a culture system comprising basic medium and knockout serum replacement (KOSR). The methods may also be applicable for selective spontaneous or directed differentiation of SCs into a selected population of somatic cells from a culture system of SCs in suspension, the method further comprising incubating said undifferentiated SCs in culture system that support respectively, spontaneous or directed differentiation of SCs into the selected population of somatic cells. The present application also discloses a culture system for expansion of stem cells (SCs) comprising a suspension of undifferentiated stem cells within basic medium and knockout serum replacement (KOSR). The methods and culture system of the invention may be used for large scale production of differentiated cells.
대표청구항▼
1. A method of expanding human pluripotent stem cells (SCs), the method comprising incubating human pluripotent SCs in suspension within a culture system comprising basic medium, a knockout serum replacement (KOSR) and a TGFβ superfamily factor,wherein the human pluripotent SCs do not undergo differ
1. A method of expanding human pluripotent stem cells (SCs), the method comprising incubating human pluripotent SCs in suspension within a culture system comprising basic medium, a knockout serum replacement (KOSR) and a TGFβ superfamily factor,wherein the human pluripotent SCs do not undergo differentiation during the expansion. 2. The method of claim 1, wherein the human pluripotent SCs are derived from human pluripotent SC colonies cultivated on a feeder layer or in a feeder free adherent culture system. 3. The method of claim 1, wherein the suspension of human pluripotent SCs comprises free floating cells, free floating clusters of cells or free floating aggregates of cells. 4. The method of claim 1, wherein the basic medium has the following composition: Concentration in uMCaCl3 (anhydrous)1800FE(N3O)39H2O0.2KCl5360MgCl3 (anhydrous)812NaCl76000NaHCO3880NaH2PO4H2O900ZnSO47H2O0.67D-glucose25000phenol red23MOPS10000sodium pyruvate230L-alanine20L-arginine HCl400L-asparagine H2O5L-cysteine10L-glutamine500Glycine400L-histidine HCl H2O200L-isoleucine800L-leucine800L-lysine HCl5L-methionine200L-phenylalanine400L-proline67L-serine400L-threonine800L-tryptophan80L-tyrosine400L-valine800D-Ca pantothenate8choline chloride80folic acid8i-inositol40niacinamide30pyridoxal HCl20riboflavin1thiamine HCl10vitamin B120.2. 5. The method of claim 1, wherein the culture system further comprises one or more element selected from the group consisting of a member of the FGF family, an extracellular matrix (ECM) component, an antibacterial agent, a non-essential amino acid, a neurotrophin, nicotinamide (NA), a bone morphogenic protein (BMP) antagonist and a serum free medium supplement. 6. The method of claim 5, wherein the ECM is selected from the group consisting of fibronectin, laminin and gelatin; the antibacterial agent is selected from the group consisting of penicillin and streptomycin; the TGFβ superfamily factor is activin A; the BMP antagonist is selected from the group consisting of noggin, chordin and gremlin; the neurotrophin is selected from the group consisting of BDNF, NT3 and NT4; and the serum free medium supplement is Nutridoma-CS. 7. The method of claim 1, wherein the human pluripotent SCs comprise human embryonic stem cells (hESCs). 8. The method of claim 1, wherein the expanding is effected in a bioreactor. 9. A culture system comprising a suspension of human pluripotent stem cells (SCs) in a culture medium comprising a basic medium, knockout serum replacement (KOSR), and a TGFβ superfamily factor. 10. The culture system of claim 9 wherein the suspension of human pluripotent stem cells comprises free floating cells, free floating clusters of cells or free floating aggregates of cells. 11. The culture system of claim 9, wherein the basic medium has the following composition: Concentration in uMCaCl3 (anhydrous)1800FE(N3O)39H2O0.2KCl5360MgCl3 (anhydrous)812NaCl76000NaHCO3880NaH2PO4H2O900ZnSO47H2O0.67D-glucose25000phenol red23MOPS10000sodium pyruvate230L-alanine20L-arginine HCl400L-asparagine H2O5L-cysteine10L-glutamine500Glycine400L-histidine HCl H2O200L-isoleucine800L-leucine800L-lysine HCl5L-methionine200L-phenylalanine400L-proline67L-serine400L-threonine800L-tryptophan80L-tyrosine400L-valine800D-Ca pantothenate8choline chloride80folic acid8i-inositol40niacinamide30pyridoxal HCl20riboflavin1thiamine HCl10vitamin B120.2. 12. The culture system of claim 9, further comprising one or more element selected from the group consisting of a member of FGF family, an extracellular matrix (ECM) component, an antibacterial agent, a non-essential amino acid, a neurotrophin, nicotinamide (NA), a bone morphogenic protein (BMP) antagonist and a serum free medium supplement. 13. The culture system of claim 12, wherein the FGF member is FGF-2, the ECM is selected from the group consisting of fibronectin, laminin and gelatin; the antibacterial agent is selected from the group consisting of penicillin and streptomycin; the TGFβ superfamily factor is activin A; the BMP antagonist is selected from the group consisting of noggin, chordin and gremlin; the neurotrophin is selected from the group consisting of BDNF, NT3, and NT4; and the serum free medium supplement is Nutridoma-CS. 14. The culture system of claim 9, wherein the human pluripotent SCs are pluripotent, human embryonic SCs (hESCs). 15. The culture system of claim 9, being free of laminin. 16. The culture system of claim 9, wherein the human pluripotent SCs are cultured in a bioreactor. 17. The method of claim 1, wherein the TGFβ superfamily factor is activin A. 18. The method of claim 1, wherein the culture system further comprises a member of the FGF family. 19. The method of claim 18, wherein the member of the FGF family factor is FGF-2. 20. A culture system comprising a suspension of human pluripotent stem cells (SCs) within in a culture medium comprising a basic medium, knockout serum replacement (KOSR), a TGFβ superfamily member, and one or more element selected from the group consisting of a member of the FGF family, an extracellular matrix (ECM) component, an antibacterial agent, a non-essential amino acid, a neurotrophin, nicotinamide (NA), a bone morphogenic protein (BMP) antagonist and a serum free medium supplement.
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이 특허에 인용된 특허 (4)
Brewer Gregory J. ; Price Paul J., Cultural medium for maintaining neural cells in ambient atmosphere.
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