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AssigneeTechnion Research & Development Foundation Limited
InventorsMichal Amit, Joseph Itskovitz-Eldor
Estimating the term…
Provided are novel serum-free culture media which comprise basic fibroblast growth factor (bFGF), transforming growth factor beta-3 and ascorbic acid at a concentration of at least about 50 microgram/ml; ascorbic acid at a concentration range of about 400-600 microgram/ml, bFGF at a concentration range of about 50-200 ng/ml, xeno-free serum replacement and a lipid mixture; the IL6RIL6 chimera at a concentration range of about 50-200 picogram per milliliter (pg/ml); or leukemia inhibitory factor (LIF) at a concentration of at least 2000 units/ml; cell cultures comprising same with pluripotent stem cells such as human embryonic stem cells and induced pluripotent stem (iPS) cells, and methods of using same for expanding pluripotent stem cells in an undifferentiated state using two-dimensional or three-dimensional culture systems; and methods of expanding iPS cells in a suspension culture devoid of substrate adherence and cell encapsulation.
A method of expanding and maintaining pluripotent stem cells in an undifferentiated state, the method comprising culturing the pluripotent stem cells in a suspension culture devoid of substrate adherence or external matrix components in a culture medium which comprises a differentiation inhibitory factor and serum replacement, said differentiation inhibitory factor consists of basic fibroblast growth factor (bFGF) at a concentration range of about 50-200 ng/ml, thereby expanding and maintaining the pluripotent stem cells in the undifferentiated state.
A method of expanding and maintaining pluripotent stem cells in an undifferentiated state, the method comprising culturing the pluripotent stem cells in a suspension culture devoid of substrate adherence or external matrix components in a culture medium being serum-free, feeder-free, matrix-free and protein carrier-free and comprising a differentiation inhibitory factor and serum replacement, wherein said differentiation inhibitory factor consists of basic fibroblast growth factor (bFGF) at a concentration range of about 50-200 ng/ml wherein the culture medium is capable of maintaining pluripotent stem cells in an undifferentiated state.
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