Simplified basic media for human pluripotent cell culture

• US 9,644,186 B2
• Filed: 03/07/2016
• Issued: 05/09/2017
• Est. Priority Date: 08/05/2010
• Status: Active Grant

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Chen, Guokai, Thomson, James A.
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US15/063,046
• Publication Number:
  US 20160194611A1
• Time in Patent Office: 428 Days
• CPC Class Codes: C12N 2500/02 Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2500/98 Xeno-free medium and culture conditions
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 2501/845 Gamma amino butyric acid [GABA]
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• 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 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

Fully defined media that support pluripotent cell viability, proliferation, cloning, and derivation, as well as methods and compositions including these media are described. Methods for deriving iPS cells from adult individuals under defined, xeno-free conditions are also described.

Claims

1. A method for deriving an induced pluripotent stem (iPS) cell under defined conditions, the method comprising the step of:

introducing reprogramming factors into a somatic cell in a defined, albumin-free medium that is free of any component obtained from a non-human animal, the medium comprising water, salts, amino acids, vitamins, glucose, an FGF, selenium, transferrin, one of insulin, IGF1, and IGF2, and one of TGF-β and

NODAL, each in an amount sufficient to support reprogramming of the somatic cell to derive an iPS cell.

2. The method of claim 1, wherein the IGF1 is Long-IGF1.

3. The method of claim 1, wherein the reprogramming step comprises contacting the cell with TGFβ

for 5-10 days.

4. The method of claim 1, further comprising the steps of:

removing the TGFβ

after 5-10 days; and

contacting the cells with a medium consisting essentially of;

water, salts, amino acids, vitamins, glucose, an FGF, selenium, one of insulin, IGF1, and IGF2, and transferrin.

5. The method of claim 1, wherein the reprogramming step comprises contacting the cell with hydrocortisone.

6. The method of claim 1, wherein the reprogramming step comprises contacting the cell with butyrate.

7. The method of claim 1, wherein the somatic cell is an adult somatic cell.

8. The method of claim 1, wherein the reprogramming step comprises introducing a viral vector into the somatic cell.

9. The method of claim 1, wherein the reprogramming step comprises introducing an episomal vector into the somatic cell.