Amplification of cell populations from embryonic stem cells
First Claim
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1. A method of culturing embryonic stem cells, comprising:
- providing a plurality of embryonic stem cells (ESC);
seeding the ESC on a substrate or suspending them in a culture medium as a single cell culture; and
incubating the seeded or suspended ESC under conditions in which they can differentiate to obtain a population of cells, wherein the ESC are not stimulated to form embryoid bodies, and wherein the proportion of a predetermined differentiated cell type in the population of cells is at least 2 times greater than the proportion of the predetermined differentiated cell type in a population obtained in the same manner except that the ESC are stimulated to form embryoid bodies.
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Abstract
Culturing embryonic stem cells without the use of embryoid bodies leads to a increase in the frequency of predetermined cell types.
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Citations
83 Claims
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1. A method of culturing embryonic stem cells, comprising:
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providing a plurality of embryonic stem cells (ESC);
seeding the ESC on a substrate or suspending them in a culture medium as a single cell culture; and
incubating the seeded or suspended ESC under conditions in which they can differentiate to obtain a population of cells, wherein the ESC are not stimulated to form embryoid bodies, and wherein the proportion of a predetermined differentiated cell type in the population of cells is at least 2 times greater than the proportion of the predetermined differentiated cell type in a population obtained in the same manner except that the ESC are stimulated to form embryoid bodies. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 67)
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- 30. A single cell population of undifferentiated embryonic stem cells (ESC) in contact with a growth factor or cytokine.
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40. A method of culturing embryonic stem cells, comprising:
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providing a plurality of embryonic stem cells (ESC);
seeding the ESC on a substrate or suspending them in a culture medium as a single cell culture; and
incubating the seeded or suspended ESC under conditions in which they can differentiate to obtain a population of cells, wherein the ESC are not stimulated to form embryoid bodies, and wherein at least a portion of the cells express a predetermined phenotype during the step of incubating, and wherein the elapsed time (“
the first elapsed time”
) before the predetermined phenotype is expressed is less than an elapsed time (“
the second elapsed time”
) for a population of cells obtained in the same manner except that the ESC are stimulated to form embryoid bodies. - View Dependent Claims (41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66)
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68. A method of obtaining a population of osteogenic cells, comprising:
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providing a population of embryonic stem cells (ESC);
seeding the ESC on a substrate as a single cell culture; and
incubating the seeded ESC in the presence of an osteogenic differentiation factor, wherein the ESC are not stimulated to form embryoid bodies. - View Dependent Claims (69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83)
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Specification