Pluripotential embryonic stem cells and methods of making same

• US 5,453,357 A
• Filed: 10/08/1992
• Issued: 09/26/1995
• Est. Priority Date: 10/08/1992
• Status: Expired due to Term

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Hogan, Brigid L. M.
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US07/958,562
• Time in Patent Office: 1,083 Days
• Field of Search: 435/7.21, 435/29, 435/240.2, 435/240.21, 435/240.3, 435/21, 435/34
• US Class Current: 435/7.21
• 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 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/04 from germ cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The present invention provides a non-mouse pluripotential embryonic stem cell which can:

(a) be maintained on feeder layers for at least 20 passages; and

(b) give rise to embryoid bodies and multiple differentiated cell phenotypes in monolayer culture. The invention further provides a method of making a pluripotential embryonic stem cell comprising administering a growth enhancing amount of basic fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor to primordial germ cells under cell growth conditions, thereby making a pluripotential embryonic stem cell.

Claims

1. A composition comprising:

(a) pluripotential embryonic stem cells; and

(b) a fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor in amounts to enhance the growth of and allow the continued proliferation of the cells.

2. The composition of claim 1, wherein the fibroblast growth factor is basic fibroblast growth factor.

3. A composition comprising:

(a) primordial germ cells; and

(b) a fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor and soluble steel factor in amounts to enhance the growth of and allow the continued proliferation of the cells.

4. The composition of claim 3, wherein the fibroblast growth factor is basic fibroblast growth factor.

5. A composition comprising:

(a) embryonic ectoderm cells; and

(b) fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor and soluble steel factor in amounts to enhance the growth of and allow the continued proliferation of the cells.

6. A composition comprising a fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor in amounts to enhance the growth of and allow the continued proliferation of primordial germ cells.

7. The composition of claim 6, wherein the fibroblast growth factor is basic fibroblast growth factor.

8. A method of making a pluripotential embryonic stem cell comprising administering a growth enhancing amount of basic fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor to primordial germ cells under cell growth conditions, thereby making a pluripotential embryonic stem cell.

9. The method of claim 8 wherein the stem cell is derived from a mammal.

10. The method of claim 9, wherein the mammal is a mouse.

11. A method of making a pluripotential embryonic stem cell comprising administering a growth enhancing amount of basic fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor to embryonic ectoderm cells under cell growth conditions, thereby making a pluripotential embryonic stem cell.

12. The method of claim 11, wherein the stem cell is derived from a mammal.

13. A method of screening cells which can be promoted to become pluripotent embryonic stem cells comprising contacting the cells with basic fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor in amounts to enhance the growth of and allow proliferation of the cells and determining which cells become pluripotent embryonic stem cells.

14. A composition comprising a fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor in amounts to promote the formation of pluripotent embryonic stem cells from primordial germ cells.