Method of enriching a mammalian cell population for mesoderm cells

• US 7,955,849 B2
• Filed: 03/24/2008
• Issued: 06/07/2011
• Est. Priority Date: 05/17/2002
• Status: Expired due to Term

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG), Mount Sinai School of Medicine (Sinai Health System)
• Original Assignee: Mount Sinai School of Medicine (Sinai Health System)
• Inventors: Keller, Gordon M., Fehling, Hans Joerg, Kubo, Atsushi, Kouskoff, Valerie
• Primary Examiner(s): Wilson; Michael C.
• Application Number:
  US12/077,976
• Publication Number:
  US 20080226558A1
• Time in Patent Office: 1,170 Days
• Field of Search: 435/455, 435/325
• US Class Current: 435/325
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• 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: G01N 33/5073 Stem cells

Abstract

The present invention provides cell populations that are enriched for mesendoderm and mesoderm, and cell populations that are enriched for endoderm. The cell populations of the invention are useful for generating cells for cell replacement therapy.

Claims

1. A method of isolating a mammalian cell population enriched for mesoderm cells comprising culturing mammalian embryoid bodies in the absence of serum and in the presence of activin at a concentration of 30 ng/ml or less, and isolating a cell population enriched for mesoderm cells after said culturing.

2. A method of generating a mammalian cell population enriched for mesoderm cells comprising:

a) culturing mammalian embryonic stem cells in differentiation culture that does not contain activin until embryoid bodies are formed, and

b) adding a serum free medium and activin at a concentration of 30 ng/ml or less to said embryoid bodies such that a cell population enriched for mesoderm is generated.