Abstract
To date, there is no suitable in vitro model to study human adult cardiac cell biology. Although embryonic stem cells are able to differentiate into cardiomyocytes in vitro, the efficiency of this process is very low. Other methods to differentiate progenitor cells into beating cardiomyocytes rely on coculturing with rat neonatal cardiomyocytes, making it difficult to study human cardiomyocyte differentiation and (patho)physiology. Here we have developed a method for efficient isolation and expansion of human cardiomyocyte progenitor cells (CMPCs) from cardiac surgical waste or alternatively from fetal heart tissue. Furthermore, we provide a detailed in vitro protocol for efficient differentiation of CMPCs into cardiomyocytes with great efficiency (80–90% of differentiation). Once CMPCs are rapidly dividing (∼1 month after isolation), differentiation can be achieved in 3–4 weeks.
This is a preview of subscription content, access via your institution
Relevant articles
Open Access articles citing this article.
-
The impact of autophagy modulation on phenotype and survival of cardiac stromal cells under metabolic stress
Cell Death Discovery Open Access 01 April 2022
-
Microengineered platforms for characterizing the contractile function of in vitro cardiac models
Microsystems & Nanoengineering Open Access 28 February 2022
-
In vitro CSC-derived cardiomyocytes exhibit the typical microRNA-mRNA blueprint of endogenous cardiomyocytes
Communications Biology Open Access 30 September 2021
Access options
Subscribe to this journal
Receive 12 print issues and online access
214,86 € per year
only 17,91 € per issue
Rent or buy this article
Get just this article for as long as you need it
$39.95
Prices may be subject to local taxes which are calculated during checkout
References
Smits, A.M. et al. The role of stem cells in cardiac regeneration. J. Cell Mol. Med. 9, 25–36 (2005).
van Vliet, P., Sluijter, J.P., Doevendans, P.A. & Goumans, M.J. Isolation and expansion of resident cardiac progenitor cells. Expert. Rev. Cardiovasc. Ther. 5, 33–43 (2007).
Mitcheson, J.S., Hancox, J.C. & Levi, A.J. Cultured adult cardiac myocytes: future applications, culture methods, morphological and electrophysiological properties. Cardiovasc. Res. 39, 280–300 (1998).
Passier, R. & Mummery, C. Cardiomyocyte differentiation from embryonic and adult stem cells. Curr. Opin. Biotechnol. 16, 498–502 (2005).
Passier, R. et al. Increased cardiomyocyte differentiation from human embryonic stem cells in serum-free cultures. Stem Cells 23, 772–780 (2005).
Braam, S.R. et al. Improved genetic manipulation of human embryonic stem cells. Nat. Methods 5, 389–392 (2008).
Goumans, M.J. et al. TGFb1 induces efficient differentiation of human cardiomyocyte progenitor cells into functional cardiomyocytes in vitro . Stem Cell Res. 1, 138–149 (2008).
van Vliet, P. et al. Progenitor cells isolated from the human heart: a potential cell source for regenerative therapy. Neth. Heart J. 16, 163–169 (2008).
Messina, E. et al. Isolation and expansion of adult cardiac stem cells from human and murine heart. Circ. Res. 95, 911–921 (2004).
Bearzi, C. et al. Human cardiac stem cells. Proc. Natl. Acad. Sci. USA 104, 14068–14073 (2007).
Urbanek, K. et al. Intense myocyte formation from cardiac stem cells in human cardiac hypertrophy. Proc. Natl. Acad. Sci. USA 100, 10440–10445 (2003).
Martin, C.M. et al. Persistent expression of the ATP-binding cassette transporter, Abcg2, identifies cardiac SP cells in the developing and adult heart. Dev. Biol. 265, 262–275 (2004).
Laugwitz, K.L. et al. Postnatal isl1+ cardioblasts enter fully differentiated cardiomyocyte lineages. Nature 433, 647–653 (2005).
Barile, L., Messina, E., Giacomello, A. & Marban, E. Endogenous cardiac stem cells. Prog. Cardiovasc. Dis. 50, 31–48 (2007).
Choi, S.C. et al. 5-Azacytidine induces cardiac differentiation of P19 embryonic stem cells. Exp. Mol. Med. 36, 515–523 (2004).
Makino, S. et al. Cardiomyocytes can be generated from marrow stromal cells in vitro . J. Clin. Invest. 103, 697–705 (1999).
Duda, D.G., Cohen, K.S., Scadden, D.T. & Jain, R.K. A protocol for phenotypic detection and enumeration of circulating endothelial cells and circulating progenitor cells in human blood. Nat. Protoc. 2, 805–810 (2007).
Nolan, T., Hands, R.E. & Bustin, S.A. Quantification of mRNA using real-time RT-PCR. Nat. Protoc. 1, 1559–1582 (2006).
Acknowledgements
This work was supported by a VIDI-grant (016.056.319, M.-J.G.) from The Netherlands Organization for Scientific Research (NWO), the Van Ruyven Foundation and BSIK program 'Dutch Program for Tissue Engineering.'
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Smits, A., van Vliet, P., Metz, C. et al. Human cardiomyocyte progenitor cells differentiate into functional mature cardiomyocytes: an in vitro model for studying human cardiac physiology and pathophysiology. Nat Protoc 4, 232–243 (2009). https://doi.org/10.1038/nprot.2008.229
Published:
Issue Date:
DOI: https://doi.org/10.1038/nprot.2008.229
This article is cited by
-
The impact of autophagy modulation on phenotype and survival of cardiac stromal cells under metabolic stress
Cell Death Discovery (2022)
-
Microengineered platforms for characterizing the contractile function of in vitro cardiac models
Microsystems & Nanoengineering (2022)
-
Genome-wide methylome pattern predictive network analysis reveal mesenchymal stem cell’s propensity to undergo cardiovascular lineage
3 Biotech (2022)
-
In vitro CSC-derived cardiomyocytes exhibit the typical microRNA-mRNA blueprint of endogenous cardiomyocytes
Communications Biology (2021)
-
Dysregulation of Notch signaling in cardiac mesenchymal cells of patients with tetralogy of Fallot
Pediatric Research (2020)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
