Differentiation of human adipose-derived stem cells into beating cardiomyocytes

J Cell Mol Med. 2010 Apr;14(4):878-89. doi: 10.1111/j.1582-4934.2010.01009.x. Epub 2010 Jan 11.


Human adipose-derived stem cells (ASCs) may differentiate into cardiomyocytes and this provides a source of donor cells for tissue engineering. In this study, we evaluated cardiomyogenic differentiation protocols using a DNA demethylating agent 5-azacytidine (5-aza), a modified cardiomyogenic medium (MCM), a histone deacetylase inhibitor trichostatin A (TSA) and co-culture with neonatal rat cardiomyocytes. 5-aza treatment reduced both cardiac actin and TropT mRNA expression. Incubation in MCM only slightly increased gene expression (1.5- to 1.9-fold) and the number of cells co-expressing nkx2.5/sarcomeric alpha-actin (27.2% versus 0.2% in control). TSA treatment increased cardiac actin mRNA expression 11-fold after 1 week, which could be sustained for 2 weeks by culturing cells in cardiomyocyte culture medium. TSA-treated cells also stained positively for cardiac myosin heavy chain, alpha-actin, TropI and connexin43; however, none of these treatments produced beating cells. ASCs in non-contact co-culture showed no cardiac differentiation; however, ASCs co-cultured in direct contact co-culture exhibited a time-dependent increase in cardiac actin mRNA expression (up to 33-fold) between days 3 and 14. Immunocytochemistry revealed co-expression of GATA4 and Nkx2.5, alpha-actin, TropI and cardiac myosin heavy chain in CM-DiI labelled ASCs. Most importantly, many of these cells showed spontaneous contractions accompanied by calcium transients in culture. Human ASC (hASC) showed synchronous Ca(2+) transient and contraction synchronous with surrounding rat cardiomyocytes (106 beats/min.). Gap junctions also formed between them as observed by dye transfer. In conclusion, cell-to-cell interaction was identified as a key inducer for cardiomyogenic differentiation of hASCs. This method was optimized by co-culture with contracting cardiomyocytes and provides a potential cardiac differentiation system to progress applications for cardiac cell therapy or tissue engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Actins / metabolism
  • Adipose Tissue / cytology*
  • Adult
  • Animals
  • Animals, Newborn
  • Azacitidine / pharmacology
  • Calcium / metabolism
  • Cell Communication / drug effects
  • Cell Differentiation* / drug effects
  • Cells, Cultured
  • Coculture Techniques
  • Culture Media / pharmacology
  • Gap Junctions / drug effects
  • Gap Junctions / metabolism
  • Gene Expression Regulation / drug effects
  • Heart / drug effects
  • Heart / physiology*
  • Humans
  • Hydroxamic Acids / pharmacology
  • Imaging, Three-Dimensional
  • Middle Aged
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Stem Cells / metabolism
  • Troponin T / genetics
  • Troponin T / metabolism


  • Actins
  • Culture Media
  • Hydroxamic Acids
  • RNA, Messenger
  • Troponin T
  • trichostatin A
  • Azacitidine
  • Calcium