Human embryonic germ cell line and methods of use
AssigneeThe Johns Hopkins University School of Medicine
InventorsJohn D. Gearhart, Michael Joseph Shamblott
Life of this patent
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Abstract
Primordial germ cells isolated from human embryonic tissue, such as from the gonadal ridges of human embryo, are disclosed. The primordial germ cells are cultured resulting in cells that resemble embryonic stem cells or embryonic germ 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.
Claims36
A method for producing human pluripotent embryonic germ (hEG) cells, comprising culturing human primordial germ cells (PGCs) in a culture medium comprising:
- (a) a ligand which binds to a receptor which can associate with glycoprotein 130 (gp 130);
- and
- (b) a growth factor;
- until cells with the morphology of human pluripotent embryonic germ cells are observed, and wherein the cells exhibit the following culture characteristics during maintenance:
- (i) dependence on a ligand that binds to a receptor, which can associate with gp 130;
- and (ii) dependence on a growth factor.
A method of maintaining a culture of human pluripotent embryonic germ (hEG) cells in a substantially undifferentiated state, wherein the cells are characterized as requiring a ligand which binds to a receptor which can heterodimerize with glycoprotein 130 (gp130) and a growth factor, the method comprising passaging the hEG cells in a culture medium comprising:
- (a) a ligand which binds to a receptor which can associate with gp130;
- and
- (b) a growth factor.
A method for producing and maintaining human pluripotent embryonic germ (hEG) cells, comprising culturing human primordial germ cells (PGCs) in a culture medium comprising:
- (a) a ligand which binds to a receptor which can associate with glycoprotein 130 (gp 130);
- and
- (b) a growth factor;
- until cells with the morphology of human pluripotent embryonic germ cells are observed, and wherein the cells exhibit the following culture characteristics during maintenance:
- (i) dependence on a ligand that binds to a receptor which can associate with gp 130;
- and
- (ii) dependence on a growth factor;
- and maintaining the hEG cells in a substantially undifferentiated state by culturing the cells in a medium containing a factor that elevates cellular cAMP levels, a growth factor, and a factor that stimulates gp 130.
A method for producing human pluripotent embryonic germ (hEG) cells, comprising culturing human primordial germ cells (PGCs) in a culture medium comprising:
- (a) an agent that binds to and activates gp130;
- and
- (b) a growth factor;
- until cells with the morphology of human pluripotent embryonic germ cells are observed, and wherein the cells exhibit the following culture characteristics during maintenance:
- (i) dependence on an agent that binds to and activates gp130;
- (ii) dependence on a growth factor.
A method of maintaining a culture of human pluripotent embryonic germ (hEG) cells in a substantially undifferentiated state, wherein the cells are characterized as requiring an agent that binds to and activates gp130 and a growth factor, the method comprising passaging the hEG cells in a culture medium comprising:
- (a) an agent that binds to and activates gp130;
- and
- (b) a growth factor.
Dependencies are inferred from the claim text.