Directed differentiation of embryonic cells
First Claim
1. A method of mapping a pathway of differentiation of a population of embryonic cells, comprising:
- (i) selecting;
(a) a set of gene expression products, wherein each gene expression product in the set is characteristic of a cell type that has undergone differentiation, such that a plurality of differentiated cell types are represented in the set; and
(b) an exogenous factor from a library of exogenous factors;
(ii) applying the exogenous factor to the population of embryonic cells;
(iii) characterizing the effect of the exogenous factor on the differentiation pathway of the population of cells by determining gene expression products in the set; and
(iv) mapping the pathway of differentiation of the cells.
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Abstract
Methods are described for mapping a pathway of differentiation of a population of embryonic cells which includes exposing the cells to an exogenous factor and measuring gene expression products that are characteristic of a particular cell type or lineage. Directing differentiation of human embryonic cells relies on dissociated embryoid bodies which are then exposed to one or more exogenous factors to enrich a culture for a particular cell type. The differentiated cells may be used for treating a medical condition in a human. Kits for determining differentiation pathways and screening exogenous factors for their utility in differentiation are provided.
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Citations
47 Claims
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1. A method of mapping a pathway of differentiation of a population of embryonic cells, comprising:
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(i) selecting;
(a) a set of gene expression products, wherein each gene expression product in the set is characteristic of a cell type that has undergone differentiation, such that a plurality of differentiated cell types are represented in the set; and
(b) an exogenous factor from a library of exogenous factors;
(ii) applying the exogenous factor to the population of embryonic cells;
(iii) characterizing the effect of the exogenous factor on the differentiation pathway of the population of cells by determining gene expression products in the set; and
(iv) mapping the pathway of differentiation of the cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 17)
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8. A method of directing differentiation of human embryonic cells to a specific cell type, comprising:
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a. permitting a population of embryonic stem cells to form embryoid bodies;
b. dissociating the embryoid bodies to provide embryonic cells for differentiating in the presence of at least one exogenous factor for an effective period of time; and
c. causing directed differentiation of human embryonic cells to form the specific cell type - View Dependent Claims (9, 10, 11, 12, 13, 14, 15, 16, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 39, 40, 43, 44, 45, 46, 47)
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18. A method of treating a subject suffering from a condition associated with degeneration of cells or malfunction of cells, comprising:
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a. accessing human embryoid body dissociated cells;
b. treating the cells with an exogenous factor;
c. causing the cells to differentiate; and
d. placing an effective amount of differentiated cells into the subject to treat the condition.
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38. A kit for determining differentiation pathways, comprising:
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(a) a plurality of sets of cell specific markers forming a panel in an assay format, the assay format including reagents for detecting the cell specific markers, the cell specific markers including a first set of markers that are characteristic of each of the ectoderm, mesoderm and endoderm of the embryo, and a second set of markers that are characteristic of a body tissue, the second set containing more than one marker; and
(b) means for detecting reagents bound to cell specific markers.
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41. A method for screening an exogenous factor to determine whether the factor is capable of causing directed differentiation in a population of human embryonic cells, comprising:
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(a) subjecting the population of cells to the exogenous growth factor;
(b) measuring the expression of receptors, the receptors being of a type that characterizes a particular differentiated cell population; and
(c) determining whether the exogenous factor enhances differentiation, maintains differentiation at a normal level or inhibits differentiation of the cell population.
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42. A panel of cell type differentiation determining markers, comprising:
- a set of reagents for specifically detecting gene expression of a plurality of ectodermal specific proteins, a plurality of mesodermal specific proteins and a plurality of endodermal specific proteins.
Specification