Alternative Compositions and Methods for the Culture of Stem Cells
First Claim
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7. The human pluripotent embryonic stem cell culture of claim 22, wherein the bone marrow stromal cell is ATCC Deposit Number CRL-11882.
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Abstract
Methods and cell culture medium for the generation and maintenance of human pluripotent embryonic stem cells are disclosed. Human embryonic stem cells are cultured with human feeder cell conditioned medium, and the embryonic stem cells maintain their pluripotent phenotype. The human pluripotent embryonic stem cells can be cultured without feeder cells, and in the presence of supplemental growth factors.
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Citations
20 Claims
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7. The human pluripotent embryonic stem cell culture of claim 22, wherein the bone marrow stromal cell is ATCC Deposit Number CRL-11882.
- 11. A method of maintaining a human pluripotent embryonic stem cell culture in an undifferentiated state for at least two passages, comprising culturing a human embryonic stem cell in a human feeder cell conditioned medium.
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13-1. The method of claim 11, comprising the steps of:
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a) isolating cells from the inner cell mass of a blastocyst; b) plating the inner cell mass cells, wherein inner cell mass-derived cell masses are formed; c) re-plating the human embryonic stem cell colony in the absence of a feeder cell; and d) adding a human feeder cell conditioned medium; thereby maintaining a human pluripotent embryonic stem cell in an undifferentiated state for at least two passages.
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17. A method of isolating and maintaining a human pluripotent embryonic stem cell culture, comprising the steps of:
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a) isolating cells from the inner cell mass of a blastocyst; b) plating the inner cell mass cells on a human feeder cell, wherein inner cell mass-derived cell masses are formed; c) re-plating the colony in the absence of a feeder cell; and d) adding a human feeder cell conditioned medium to thereby isolate and maintain a human pluripotent embryonic stem cell in an undifferentiated state for at least two passages. - View Dependent Claims (18, 19, 20)
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