Method of in vitro differentiation of transplantable neural precursor cells from primate embryonic stem cells

• US 6,887,706 B2
• Filed: 10/03/2001
• Issued: 05/03/2005
• Est. Priority Date: 10/03/2001
• Status: Expired due to Term

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Thomson, James A., Zhang, Su-Chun, Duncan, Ian David
• Primary Examiner(s): Gozo, David
• Assistant Examiner(s): Nguyen, Quang
• Application Number:
  US09/970,382
• Publication Number:
  US 20030068819A1
• Time in Patent Office: 1,308 Days
• Field of Search: 435/325, 435/363, 435/366, 435/368, 435377-378
• US Class Current: 435/377
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/392 Sexual steroids
• CPC Class Codes: C12N 2501/91 Heparin
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

A method of differentiating embryonic stem cells into neural precursor cells is disclosed. In one embodiment the method comprises the steps of (a) obtaining an embryonic stem cell culture, (b) propagating the stem cells, (c) forming embryoid bodies from the stem cells, and (d) culturing the embryoid bodies in a medium containing an effective amount of fibroblast growth factor 2, wherein neural precursor cells will be generated and isolated.

Claims

1. A method of differentiating primate embryonic stem cells into neural precursor cells, comprising the steps of:

(a) obtaining a primate embryonic stem cell culture,

(b) propagating the stem cells, wherein embryoid bodies are formed, and

(c) culturing the embryoid bodies in a medium consisting essentially of DMEM/F12, insulin, transferrin, progesterone, putrescine, sodium selenite, heparin and an effective amount of fibroblast growth factor 2, wherein neural precursor cells are generated and wherein the neural precursor cells form rosette formations.