Chemically defined conditions for human iPSC derivation and culture

Nat Methods. 2011 May;8(5):424-9. doi: 10.1038/nmeth.1593. Epub 2011 Apr 10.

Abstract

We re-examine the individual components for human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) culture and formulate a cell culture system in which all protein reagents for liquid media, attachment surfaces and splitting are chemically defined. A major improvement is the lack of a serum albumin component, as variations in either animal- or human-sourced albumin batches have previously plagued human ESC and iPSC culture with inconsistencies. Using this new medium (E8) and vitronectin-coated surfaces, we demonstrate improved derivation efficiencies of vector-free human iPSCs with an episomal approach. This simplified E8 medium should facilitate both the research use and clinical applications of human ESCs and iPSCs and their derivatives, and should be applicable to other reprogramming methods.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biopsy
  • Cattle
  • Cell Culture Techniques / methods*
  • Cell Proliferation
  • Cell Survival
  • Coated Materials, Biocompatible
  • Culture Media / chemistry*
  • Culture Media, Serum-Free / chemistry
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Fibroblasts / cytology
  • Gene Expression
  • Growth Substances
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Karyotyping
  • Serum Albumin, Bovine
  • Skin / cytology
  • Vitronectin

Substances

  • Coated Materials, Biocompatible
  • Culture Media
  • Culture Media, Serum-Free
  • Growth Substances
  • Vitronectin
  • Serum Albumin, Bovine