METHOD FOR EFFICIENT TRANSFER OF HUMAN BLASTOCYST-DERIVED STEM CELLS (hBS CELLS) FROM A FEEDER-SUPPORTED TO A FEEDER-FREE CULTURE SYSTEM
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Abstract
Technology is provided for the transfer of human blastocyst-derived stem cells (hBS cells) to a feeder-free culture system and propagation of the cells in such a feeder-free culture system, the method comprising the following steps of (a) transferring the blastocyst-derived stem cells from feeder to feeder free culture by mechanical treatment, (b) culturing the blastocyst-derived stem cells under feeder cell free growth conditions in a suitable growth medium and/or on a suitable support substrate, and (c) optionally passaging the blastocyst derived stem cell line every 3-10 days by enzymatic and/or mechanical treatment. The application of hBS cells cultured under a feeder-free condition in medicine (e.g., myocardial regeneration) and screening and toxicity testing also is provided.
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Citations
48 Claims
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1-39. -39. (canceled)
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40. A medicament for transplantation of hBS cells into a mammal for the prevention or treatment of a disease comprising an effective concentration of hBS cells and one or more agents selected from the group consisting of therapeutically active substances, prophylactically active substances, engraftment improving agents, viability improving agents, differentiation improving agents, and immunosuppressive agents, wherein said hBS cells are formed by the method comprising:
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(a) transferring the hBS cells from feeder to feeder-free culture without enzymatic treatment but solely by mechanical treatment; and (b) following step (a), culturing the hBS cells under feeder-free growth conditions in a suitable growth medium and on a suitable support substrate, wherein said suitable growth medium is a cell-free medium conditioned by previous exposure to feeder cells, and said suitable support substrate comprises a cell-free matrix comprising extracellular matrix proteins. - View Dependent Claims (41, 42, 43, 44, 45, 46, 47, 48)
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Specification