Compositions and methods of making embryonic stem cells

• US 6,251,671 B1
• Filed: 02/28/1997
• Issued: 06/26/2001
• Est. Priority Date: 02/28/1996
• Status: Expired due to Term

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Zhao, Guang-Quan, Hogan, Brigid L. M.
• Primary Examiner(s): Saucier, Sandra E.
• Application Number:
  US08/808,346
• Time in Patent Office: 1,579 Days
• Field of Search: 435/384, 435/385, 435/386, 435/387
• US Class Current: 435/384
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A61K 38/1875 Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: A61P 15/08 for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 15/16 Masculine contraceptives
• CPC Class Codes: A61P 17/14 for baldness or alopecia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/125 Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 5/061 Sperm cells, spermatogonia
• CPC Class Codes: G01N 2510/00 Detection of programmed cell death, i.e. apoptosis
• CPC Class Codes: G01N 2800/367 Infertility, e.g. sperm disorder, ovulatory dysfunction
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5017 for testing neoplastic activity
• CPC Class Codes: G01N 33/5044 : involving specific cell types
• CPC Class Codes: G01N 33/5073 : Stem cells
• CPC Class Codes: G01N 33/689 : related to pregnancy or the gonads

Abstract

The invention relates to cell proliferation, cell differentiation, male infertility, male fertility and to compositions and methods involved therein. Also methods of culturing spermatogonial stem cells with bone morphogenetic protein 8 are disclosed.

Claims

1. A method of proliferating mammalian spermatogonial stem cells, consisting of culturing spermatogonial stem cells with a growth factor wherein said growth factor is bone morphogenetic protein 8 to effect proliferation of said cells, wherein said fragment adequately supports survival of spermatocytes in culture.

2. The method of claim 1, wherein said bone morphogenetic protein 8 is selected from the group consisting of at least one of mouse bone morphogenetic protein 8A, mouse bone morphogenetic protein 8B, human bone morphogenetic protein 8, rat bone morphogenetic protein 8, rabbit bone morphogenetic protein 8, guinea pig bone morphogenetic protein 8, goat bone morphogenetic protein 8, pig bone morphogenetic 8, bovine bone morphogenetic protein 8, and a dimer of the C-terminal TGFβ

-like portion of bone morphogenetic protein 8.

3. The method of claim 1, wherein said mammalian spermatogonial cells are selected from the group consisting of mouse, human, rat, rabbit, guinea pig, goat, pig and bovine cells.

4. A method of extending viability of a mammalian spermatogonial cell population, consisting of culturing said spermatogonial cell population with a growth factor wherein said growth factor is bone morphogenetic protein 8 wherein said fragment adequately supports survival of spermatocytes in culture, thereby extending the viability of said cultured spermatogonial cell population.

5. The method of claim 4, wherein said bone morphogenetic protein 8 is selected from the group consisting of at least one of mouse bone morphogenetic 8A, mouse bone morphogenetic protein 8B, human bone morphogenetic protein 8, rat bone morphogenetic protein 8, rabbit bone morphogenetic protein 8, guinea pig bone morphogenetic protein 8, goat bone morphogenetic protein 8, pig bone morphogenetic protein 8, bovine bone morphogenetic protein 8, and dimer of the C-terminal TGFβ

-like portion of bone morphogenetic protein 8.

6. The method of claim 4, wherein said mammalian spermatogonial cells are selected from the group consisting of mouse, human, rat rabbit, guinea pig, goat, pig and bovine cells.

7. A method of selectively obtaining a proliferating population of spermatogonial stem cells in culture, consisting of adding a growth factor wherein said growth factor is bone morphogenetic protein 8 to said population of cells, wherein said fragment adequately supports survival of spermatocytes in culture, thereby selectively obtaining a proliferating population of spermatogonial stem cells.

8. The method of claim 7, wherein said bone morphogenetic protein 8 is selected from the group consisting of at least one of mouse bone morphogenetic protein 8A, mouse bone morphogenetic protein 8B, human bone morphogenetic protein 8, rat bone morphogenetic protein 8, rabbit bone morphogenetic protein 8, guinea pig bone morphogenetic protein 8, goat bone morphogenetic protein 8, pig bone morphogenetic protein 8, bovine bone morphogenetic protein 8, and a dimer of the C-terminal TGFβ

-like portion of bone morphogenetic protein 8.

9. The method of claim 7, wherein said mammalian spermatogonial cells are selected from the group consisting of mouse, human, rat rabbit, guinea pig, goat, pig and bovine cells.