Scalable production of embryonic stem cell-derived cardiomyocytes

Peter W. Zandstra, C. Bauwens, T. Yin, Q. Liu, H. Schiller, R. Zweigerdt, K. B S Pasumarthi, Loren Field

Research output: Contribution to journalArticle

219 Citations (Scopus)

Abstract

Cardiomyocyte transplantation could offer a new approach to replace scarred, nonfunctional myocardium in a diseased heart. Clinical application of this approach would require the ability to generate large numbers of donor cells. The purpose of this study was to develop a scalable, robust, and reproducible process to derive purified cardiomyocytes from genetically engineered embryonic stem (ES) cells. ES cells transfected with a fusion gene consisting of the α-cardiac myosin heavy chain (MHC) promoter driving the aminoglycoside phosphotransferase (neomycin resistance) gene were used for cardiomyocyte enrichment. The transfected cells were aggregated into embyroid bodies (EBs), inoculated into stirred suspension cultures, and differentiated for 9 days before selection of cardiomyocytes by the addition of G418 with or without retinole acid (RA). Throughout the culture period, EB and viable cell numbers were measured. In addition, flow cytometric analysis was performed to monitor sarcomeric myosin (a marker for cardiomyocytes) and Oct-4 (a marker for undifferentiated ES cells) expression. Enrichment of cardiomyocytes was achieved in cultures treated with either G418 and retinoic acid (RA) or with G418 alone. Eighteen days after differentiation, G418-selected flasks treated with RA contained approximately twice as many cells as the nontreated flasks, as well as undetectable levels of Oct-4 expression, suggesting that RA may promote cardiac differentiation and/or survival. Immunohistological and electron microscopic analysis showed that the harvested cardiomyocytes displayed many features characteristic of native cardiomyocytes. Our results demonstrate the feasibility of large-scale production of viable, ES cell-derived cardiomyocytes for tissue engineering and/or implantation, an approach that should be transferable to other ES cell derived lineages, as well as to adult stem cells with in vitro cardiomyogenic activity.

Original languageEnglish
Pages (from-to)767-778
Number of pages12
JournalTissue Engineering
Volume9
Issue number4
DOIs
StatePublished - Aug 2003

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Embryonic Stem Cells
Stem cells
Cardiac Myocytes
Acids
Genes
Cell Count
Tissue engineering
Kanamycin Kinase
Cardiac Myosins
Adult Stem Cells
Fusion reactions
Neomycin
Myosin Heavy Chains
Gene Fusion
Cell Lineage
Tissue Engineering
Myosins
Tretinoin
Electrons
Heart Diseases

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology

Cite this

Zandstra, P. W., Bauwens, C., Yin, T., Liu, Q., Schiller, H., Zweigerdt, R., ... Field, L. (2003). Scalable production of embryonic stem cell-derived cardiomyocytes. Tissue Engineering, 9(4), 767-778. https://doi.org/10.1089/107632703768247449

Scalable production of embryonic stem cell-derived cardiomyocytes. / Zandstra, Peter W.; Bauwens, C.; Yin, T.; Liu, Q.; Schiller, H.; Zweigerdt, R.; Pasumarthi, K. B S; Field, Loren.

In: Tissue Engineering, Vol. 9, No. 4, 08.2003, p. 767-778.

Research output: Contribution to journalArticle

Zandstra, PW, Bauwens, C, Yin, T, Liu, Q, Schiller, H, Zweigerdt, R, Pasumarthi, KBS & Field, L 2003, 'Scalable production of embryonic stem cell-derived cardiomyocytes', Tissue Engineering, vol. 9, no. 4, pp. 767-778. https://doi.org/10.1089/107632703768247449
Zandstra PW, Bauwens C, Yin T, Liu Q, Schiller H, Zweigerdt R et al. Scalable production of embryonic stem cell-derived cardiomyocytes. Tissue Engineering. 2003 Aug;9(4):767-778. https://doi.org/10.1089/107632703768247449
Zandstra, Peter W. ; Bauwens, C. ; Yin, T. ; Liu, Q. ; Schiller, H. ; Zweigerdt, R. ; Pasumarthi, K. B S ; Field, Loren. / Scalable production of embryonic stem cell-derived cardiomyocytes. In: Tissue Engineering. 2003 ; Vol. 9, No. 4. pp. 767-778.
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