Pluripotential embryonic stem cells and methods of making same

• US 7,153,684 B1
• Filed: 03/06/1997
• Issued: 12/26/2006
• Est. Priority Date: 10/08/1992
• Status: Expired due to Fees

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Hogan, Brigid L. M.
• Primary Examiner(s): WOITACH, JOSEPH T
• Application Number:
  US08/813,829
• Time in Patent Office: 3,582 Days
• Field of Search: 435/235
• US Class Current: 435/325
• 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 2502/99 genetically modified cells
• 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
• CPC Class Codes: C12N 5/0611 Primordial germ cells, e.g. embryonic germ cells [EG]

Abstract

The present invention provides a non-mouse, including human, 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 culturing germ cells and germ cell progenitors in a composition comprising 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.

Also provided are compositions useful to produce the pluripotent embryonic stem cells and methods of screening associated with the method of making the embryonic stem cell.

Claims

1. An isolated human PGC-derived pluripotential stem cell which can:

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

(b) give rise to embryoid bodies and multiple differentiated cell phenotypes in monolayer culture, wherein the cell has a normal karyotype; and

(c) stains positive for SSEA-1.

2. The cell of claim 1, wherein the cell stains positive for alkaline phosphatase.

3. An isolated human PGC-derived pluripotential 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; and

(c) stains positive for SSEA-1.

4. The cell of claim 3, wherein the cell stains positive for alkaline phosphatase.