Human enbryonic germ cell and methods of use

• US 6,331,406 B1
• Filed: 12/12/1997
• Issued: 12/18/2001
• Est. Priority Date: 03/31/1997
• Status: Expired due to Fees

• Current Assignee: Johns Hopkins University
• Original Assignee: The Johns Hopkins University School of Medicine
• Inventors: Gearhart, John D., Shamblott, Michael Joseph
• Primary Examiner(s): Weber, Jon P.
• Application Number:
  US08/989,744
• Time in Patent Office: 1,467 Days
• Field of Search: 517/2, 435/375, 435/395, 435/377, 435/7.21
• US Class Current: 435/7.21
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0611 Primordial germ cells, e.g. embryonic germ cells [EG]
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/689 related to pregnancy or the gonads

Abstract

Primordial germ cells extracted from post blastocyst human embryos, such as from the gonadal ridges of a 8-11 week LMP human embryo, are disclosed. The primordial germ cells are cultured in long term culture (more than 30 days) resulting in cells that resemble embryonic stem cells in morphology and pluripotency. The cells are maintained several months in culture and can be genetically manipulated using transgenic technology to insert heterologous genetic material.

Claims

1. An in vivo method for determining the effect of a compound on a human pluripotent embryonic germ (hEG) cell function comprising:

a) incubating the compound and at least one hEG cell under conditions sufficient to allow the compound and the cell to interact, wherein the hEG cell is dependent on a growth factor and a factor which binds to a receptor which can heterodimerize with gp 130 during maintenance; and

b) determining whether the compound has an effect on cell function compared to hEG cells not treated with the compound.

2. The method of claim 1, wherein the effect is inhibition of a cell function.

3. The method of claim 1, wherein the effect is stimulation of a cell function.

4. The method of claim 1, wherein the compound is a peptide.

5. The method of claim 1, wherein the compound is a peptidomimetic.

6. The method of claim 1, wherein the at least one hEG cell is on a solid support.

7. The method of claim 6, wherein the solid support is a chip.

8. The method of claim 1, wherein the hEG cell function is selected from the group consisting of differentiation, cell proliferation, production of a protein, gene expression, production of a growth factor, response to a growth factor and modulation of cell membrane permeability.

9. The method of claim 1, wherein the hEG cell is an isolated hEG cell.

10. The method of claim 9, wherein the hEG cell is SSEA-1 positive.

11. The method of claim 1, wherein the hEG cell is SSEA-1 positive.

12. The method of claim 1, wherein the hEG cell function comprises differentiation into a restricted developmental lineage cell.

13. The method of claim 12, wherein the restricted developmental lineage cell is selected from the group consisting of a neuron, a vascular endothelial cell, a cardiomyocyte, a hematopoietic cell, and a skeletal muscle cell.

14. The method of claim 1, wherein the hEG cell is dependent on a factor that elevates cAMP.

15. The method of claim 14, wherein the factor that elevates cAMP is selected from the group consisting of forskolin, cholera toxin, isobutylmethylxanthine and dibutyladenosine cyclic monophosphate.

16. The method of claim 15, wherein the factor that elevates cAMP is forskolin.