Microscale generation of cardiospheres promotes robust enrichment of cardiomyocytes derived from human pluripotent stem cells

Stem Cell Reports. 2014 Aug 12;3(2):260-8. doi: 10.1016/j.stemcr.2014.06.002. Epub 2014 Jul 4.


Cardiomyocytes derived from human pluripotent stem cells (hPSCs) are a promising cell source for regenerative medicine, disease modeling, and drug discovery, all of which require enriched cardiomyocytes, ideally ones with mature phenotypes. However, current methods are typically performed in 2D environments that produce immature cardiomyocytes within heterogeneous populations. Here, we generated 3D aggregates of cardiomyocytes (cardiospheres) from 2D differentiation cultures of hPSCs using microscale technology and rotary orbital suspension culture. Nearly 100% of the cardiospheres showed spontaneous contractility and synchronous intracellular calcium transients. Strikingly, from starting heterogeneous populations containing ∼10%-40% cardiomyocytes, the cell population within the generated cardiospheres featured ∼80%-100% cardiomyocytes, corresponding to an enrichment factor of up to 7-fold. Furthermore, cardiomyocytes from cardiospheres exhibited enhanced structural maturation in comparison with those from a parallel 2D culture. Thus, generation of cardiospheres represents a simple and robust method for enrichment of cardiomyocytes in microtissues that have the potential use in regenerative medicine as well as other applications.

Publication types

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

MeSH terms

  • Actinin / metabolism
  • Albumins / chemistry*
  • Batch Cell Culture Techniques
  • Calcium / metabolism
  • Cell Differentiation
  • Cell Line
  • Fibroblasts / cytology
  • Homeobox Protein Nkx-2.5
  • Homeodomain Proteins / metabolism
  • Humans
  • Muscle Contraction
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Myosin Heavy Chains / metabolism
  • Pluripotent Stem Cells / cytology*
  • Polyesters / chemistry*
  • Transcription Factors / metabolism
  • Troponin I / metabolism


  • Albumins
  • Homeobox Protein Nkx-2.5
  • Homeodomain Proteins
  • NKX2-5 protein, human
  • PB-127 compound
  • Polyesters
  • Transcription Factors
  • Troponin I
  • Actinin
  • Myosin Heavy Chains
  • Calcium