Differentiation of human ES cell line KIND-2 to yield tripotent cardiovascular progenitors

  • Harsha Pawani
  • Punam Nagvenkar
  • Prasad Pethe
  • Deepa Bhartiya
Article

Abstract

Human embryonic stem cells (hESCs) have the ability to differentiate into all the three lineages and are an ideal starting material to obtain cells of desired lineage for regenerative medicine. Continued efforts are needed to evolve more robust protocols to obtain cells of desired lineages and in larger numbers. Also, it has now been realized that rather than transplanting fully committed cells differentiated in vitro, it may be ideal to transplant committed progenitors which retain the intrinsic ability to proliferate and also differentiate better into multiple lineages based on the in vivo cues. For cardiac regeneration, the desired progenitor is a multipotent cardiovascular progenitor which has the ability to regenerate cardiomyocytes, endothelial cells, and also smooth muscle cells. The present study was undertaken to carefully compare three widely used protocols to differentiate hESCs into cardiac progenitors, viz., spontaneous differentiation, differentiation by END-2-conditioned medium, and directed differentiation using growth factors followed by quantitative PCR to study the relative expression of early cardiovascular markers. hESC differentiation mimicked the early embryonic development, and the transition into mesoendoderm, mesoderm, early cardiac progenitors, and cardiac cells associated with spontaneous beating was clearly evident in all the three groups. However, compared to spontaneous and END-2-associated differentiation, directed differentiation led to several-fold higher expression of cardiac transcripts (>75-fold Nkx2.5 and >150-fold Tbx5) in response to the stage-specific addition of well-established cardiogenic inducers and inhibitors of specific signaling pathways. We propose to use tripotent cardiovascular progenitors derived by directed differentiation for further preclinical studies.

Keywords

Human embryonic stem cells Cardiomyocytes Progenitors Differentiation 

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Copyright information

© The Society for In Vitro Biology 2012

Authors and Affiliations

  • Harsha Pawani
    • 1
  • Punam Nagvenkar
    • 1
  • Prasad Pethe
    • 1
  • Deepa Bhartiya
    • 1
  1. 1.Stem Cell Biology DepartmentNational Institute for Research in Reproductive HealthMumbaiIndia

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