Differentiation of human embryonic stem cells to cardiomyocytes on microcarrier cultures

Sherwin Ting, Marti Lecina, Shaul Reuveny, Steve Oh

Research output: Indexed journal article Articlepeer-review

10 Citations (Scopus)

Abstract

We have developed an improved cardiomyocyte differentiation protocol where we stabilized embryoid bodies (EB) in serum- and insulin-free medium (bSFS) supplemented with p38 MAP kinase inhibitor (SB203580) by addition of 10 μm laminin-coated positively charged (protamine sulfate derivatized TSKgel Tresyl-5PW) microcarriers. This protocol achieved a maximum 3-fold cell expansion, differentiation efficiency of 20%, and an overall cardiomyocyte yield of 3 × 105 CM/ml in static conditions. In comparison, EB cultures achieved 1.5-fold cell expansion, differentiation efficiency of 15%, and an overall cardiomyocyte yield of 1.1 × 105 CM/ml. The scalability of this platform was demonstrated in suspended spinner cultures, producing a maximum of 2.14 × 105 CM/ml in 50-ml cultures. This yield is two-fold higher than the control static EB-based platform (1.1 × 105 CM/ml), and seven-fold higher than yields reported in literature, 3.1-9 × 104 CM/ml. The robustness of this protocol was tested with HES-3 and H1 cell lines.

Original languageEnglish
Article number1D.7
JournalCurrent Protocols in Stem Cell Biology
Volume21
Issue number1
DOIs
Publication statusPublished - 1 May 2012
Externally publishedYes

Keywords

  • Cardiomyocytes;SB203580
  • Hesc;differentiation
  • Human embryonic
  • Microcarriers
  • Scale up
  • Stem cells

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