[특허]Micro-carrier culture system for rapid expansion of human embryonic stem cells

Micro-carrier culture system for rapid expansion of human embryonic stem cells 원문보기

IPC분류정보
국가/구분	United States(US) Patent 등록
국제특허분류(IPC7판)	C12N-015/02
출원번호	US-0763884 (2010-04-20)
등록번호	US-8097458 (2012-01-17)
발명자 / 주소	Mandalam, Ramkumar Xu, Chunhui Gold, Joseph D. Carpenter, Melissa K.
출원인 / 주소	Geron Corporation
대리인 / 주소	Mittler, E. Stewart
인용정보	피인용 횟수 : 0 인용 특허 : 45

초록 ▼

This disclosure provides an improved system for culturing human pluripotent stem cells. Traditionally, pluripotent stem cells are cultured on a layer of feeder cells (such as mouse embryonic fibroblasts) to prevent them from differentiating. In the system described here, the role of feeder cells is replaced by components added to the culture environment that support rapid proliferation without differentiation. Effective features are a suitable support structure for the cells, and an effective medium that can be added fresh to the culture without being preconditioned by another cell type. Culturing human embryonic stem cells in fresh medium according to this invention causes the cells to expand surprisingly rapidly, while retaining the ability to differentiate into cells representing all three embryonic germ layers. This new culture system allows for bulk proliferation of pPS cells for commercial production of important products for use in drug screening and human therapy.

대표청구항 ▼

1. A method of expanding primate pluripotent stem cells (pPS cell) in an undifferentiated state comprising a) attaching a pPS cell to a microcarrier comprising a matrix and b) providing the pPS cells with a nutrient media and c) expanding 2 fold the pPS cells in an undifferentiated state. 2. The method of claim 1, wherein the matrix comprises an extracellular matrix protein. 3. The method of claim 1, wherein the matrix comprises a soluble preparation of Engelbreth-Holm-Swarm tumor. 4. The method of claim 1, wherein the nutrient media comprises 20% fetal bovine serum. 5. The method of claim 1, wherein the nutrient media comprises 20% serum replacement. 6. The method of claim 1, wherein the nutrient media comprises albumin. 7. The method of claim 1, wherein the nutrient media comprises insulin, 8. The method of claim 1, wherein the nutrient media comprises transferrin. 9. The method of claim 1, wherein the nutrient media is a media conditioned by a cell. 10. The method of claim 1, wherein the nutrient media is non-conditioned media comprising fibroblast growth factor (FGF). 11. The method of claim 10 wherein the FGF is FGF 2. 12. The method of claim 10, wherein the FGF is FGF 4. 13. A method of expanding cells expressing the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 in an undifferentiated state comprising a) attaching the cells expressing the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 to a microcarrier comprising a matrix and b) providing the cells expressing the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 with a nutrient media and c) expanding 2 fold the cells expressing, the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 in an undifferentiated state. 14. The method of claim 13, wherein the matrix comprises an extracellular matrix protein. 15. The method of claim 13, wherein the matrix comprises a soluble preparation of Engelbreth-Holm-Swarm tumor. 16. The method of claim 13, wherein the nutrient media comprises 20% fetal bovine serum. 17. The method of claim 13, wherein the nutrient media comprises 20% serum replacement. 18. The method of claim 13, wherein the nutrient media comprises albumin. 19. The method of claim 13, wherein the nutrient media comprises insulin. 20. The method of claim 13, wherein the nutrient media comprises transferrin. 21. The method of claim 13, wherein the nutrient media is a media conditioned by a cell. 22. The method of claim 13, wherein the nutrient media is non-conditioned media comprising fibroblast growth factor (FGF). 23. The method of claim 13 wherein the FGF is FGF 2. 24. The method of claim 13, wherein the FGF is FGF 4.

이 특허에 인용된 특허 (45)

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IPC	Description
A	생활필수품
A62	인명구조; 소방(사다리 E06C)
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A62B-1/08	.. 윈치 또는 풀리에 제동기구가 있는 것

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연합인증

Micro-carrier culture system for rapid expansion of human embryonic stem cells 원문보기

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Micro-carrier culture system for rapid expansion of human embryonic stem cells 원문보기

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연구과제 타임라인

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