Method of deriving progenitor cell line
AssigneeAgency for Science, Technology and Research
InventorsSai Kiang LIM, Elias LYE
Life of this patent
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Abstract
We disclose a method comprising: (a) providing an embryonic stem (ES) cell; and (b) establishing a progenitor cell line from the embryonic stem cell; in which the progenitor cell line is selected based on its ability to self-renew. Preferably, the method selects against somatic cells based on their inability to self-renew. Preferably, the progenitor cell line is derived or established in the absence of co-culture, preferably in the absence of feeder cells, which preferably selects against embryonic stem cells. Optionally, the method comprises (d) deriving a differentiated cell from the progenitor cell line.
Claims12
A method of screening a candidate molecule for its effect on a mesenchymal progenitor cell, the method comprising:
- (a) deriving a mesenchymal progenitor cell line from a parental embryonic stem cell in the presence of a candidate molecule, wherein the mesenchymal progenitor cell line comprises mesenchymal stem cells, the method comprising:
- (i) providing the parental embryonic stem cell or descendents of the parental embryonic stem cell obtained by dispersing an embryonic stem cell colony with trypsin;
- (ii) culturing the parental embryonic stem cell in the absence of feeder cells in conditions that promote growth of putative mesenchymal progenitor cells and at least retard growth or propagation of embryonic stem cells;
- (iii) selecting mesenchymal progenitor cells which self-renew;
- and
- (iv) establishing in the absence of transformation a mesenchymal progenitor cell line from the mesenchymal progenitor cells which self-renew;
- wherein the mesenchymal progenitor cell line is maintainable in cell culture for more than 20 generations, and wherein the mesenchymal progenitor cell line is lineage restricted compared to the parental embryonic stem cell;
- and
- (b) determining the effect of a candidate molecule on mesenchymal progenitor cells treated with the candidate molecule compared with untreated mesenchymal progenitor cells or mesenchymal progenitor cells treated with an inert compound.
2-12. (canceled)
A method of screening a candidate molecule for its effect on a cell differentiated from a mesenchymal progenitor cell line, the method comprising:
- (a) deriving a mesenchymal progenitor cell line from a parental embryonic stem cell, wherein the mesenchymal progenitor cell line comprises mesenchymal stem cells, the method comprising:
- (i) providing the parental embryonic stem cell or descendents of the parental embryonic stem cell obtained by dispersing an embryonic stem cell colony with trypsin;
- (ii) culturing the parental embryonic stem cell in the absence of feeder cells in conditions that promote growth of putative mesenchymal progenitor cells and at least retard growth or propagation of embryonic stem cells;
- (iii) selecting mesenchymal progenitor cells which self-renew;
- and
- (iv) establishing in the absence of transformation a mesenchymal progenitor cell line from the mesenchymal progenitor cells which self-renew;
- wherein the mesenchymal progenitor cell line is maintainable in cell culture for more than 20 generations, and wherein the mesenchymal progenitor cell line is lineage restricted compared to the parental embryonic stem cell;
- (b) deriving a differentiated cell from the mesenchymal progenitor cell line;
- (c) treating the differentiated cell with a candidate molecule;
- and
- (d) determining the effect of a candidate molecule on the differentiated cell treated with the candidate molecule compared with untreated differentiated cells or differentiated cells treated with an inert compound.
Dependencies are inferred from the claim text.