Claims
- 1. A method to produce an immortalized cell population, comprising:
- (a) transforming an embryonic stem cell population with an immortalizing gene to create a transformed stem cell population;
- (b) culturing said transformed stem cell population under effective conditions to produce a transformed embryoid body cell population; and
- (c) incubating said transformed embryoid body cell population under conditions suitable to obtain an immortalized cell population that differentiates into cellular lineages comprising primitive erythroid cells and definitive erythroid cells.
- 2. The method of claim 1, wherein said embryonic stem cell population is derived from a mammalian embryo.
- 3. The method of claim 1, wherein said immortalizing gene comprises human HOX11.
- 4. The method of claim 1, wherein said immortalizing gene is contained in the recombinant molecule MSCV-HOX11.
- 5. The method of claim 1, wherein said step of culturing comprises culturing said transformed embryonic stem cells in an embryoid body cell medium comprising about 15% serum selected from the group consisting of platelet-poor fetal bovine serum and pre-selected normal fetal calf serum.
- 6. The method of claim 5, wherein said embryoid body cell medium does not include leukocyte inhibitory factor.
- 7. The method of claim 1, wherein said step of culturing is performed for from about 3 days to about 8 days.
- 8. The method of claim 1, wherein said step of incubating comprises culturing said embryoid body cell population in a medium comprising serum selected from the group consisting of platelet-poor fetal bovine serum and pre-selected normal fetal calf serum, and a growth factor selected from the group consisting of C-kit ligand, interleukin 3, erythropohetin, and combinations thereof.
- 9. The method of claim 1, wherein said step of incubating comprises culturing said embryoid body cell population in a medium comprising serum selected from the group consisting of platelet-poor fetal bovine serum and pre-selected normal fetal calf serum, and a combination of growth factors selected from the group consisting of C-kit ligand combined with erythropoietin and interleukin 3 combined with erythropoietin.
- 10. The method of claim 1, wherein said step of culturing is performed for about 1 day to about 7 days.
- 11. The method of claim 1, wherein said step of culturing comprises culturing said transformed stem cell population in an embryoid body cell medium comprising from about 5% to about 30% serum selected from the group consisting of platelet-poor fetal bovine serum and pre-selected normal fetal calf serum.
- 12. The method of claim 1, wherein said step of incubating comprises culturing said embryoid body cell population in a medium comprising serum selected from the group consisting of platelet-poor fetal bovine serum and pre-selected normal fetal calf serum, and C-kit ligand.
- 13. The method of claim 1, wherein said step of incubating is performed for about 2 days to about 15 days.
- 14. The method of claim 1, wherein said step of incubating is performed for about 3 days to about 6 days.
- 15. The method of claim 1, wherein said step of incubating is performed at about 37.degree. C. in an about 5% CO.sub.2 environment.
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application is a divisional application of U.S. application Ser. No. 08/570,211, filed Dec. 11, 1995, entitled "NOVEL EMBRYONIC CELL POPULATIONS AND METHODS TO ISOLATE SUCH POPULATIONS", now U.S. Pat. No. 5,874,301, issued on Feb. 23, 1999, which is a continuation-in-part of U.S. patent application Ser. No. 08/343,686, filed Nov. 21, 1994, entitled "NOVEL EMBRYONIC CELL POPULATIONS AND METHODS TO ISOLATE SUCH POPULATIONS" now U.S. Pat. No. 5,914,268. Both U.S. Pat. No. 5,874,301 and U.S. Pat. No. 5,914,268 are incorporated herein by this reference in their entirety. U.S. Pat. No. 5,874,301 is also a continuation-in-part of PCT Patent Application Serial No. PCT/US95/14495, filed Nov. 20, 1995, entitled "NOVEL EMBRYONIC CELL POPULATIONS AND METHODS TO ISOLATE SUCH POPULATIONS", which is also incorporated herein by this reference in its entirety.
GOVERNMENT RIGHTS
This invention was made in part with government support under HL 48834-02, awarded by the National Institutes of Health. The government has certain rights to this invention.
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Divisions (1)
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Number |
Date |
Country |
Parent |
570211 |
Dec 1995 |
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Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
343686 |
Nov 1994 |
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Parent |
PCTUS9514495 |
Nov 1995 |
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