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Exposure to cardiomyogenic stimuli fails to transdifferentiate human umbilical cord blood-derived mesenchymal stem cells

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Abstract

The ability of human umbilical cord blood-derived mesenchymal stem cells (UCBMSCs) to transdifferentiate towards cardiomyocytes remains unclear. The aim of this study was to direct UCBMSCs to the cardiac lineage by exposure to: (1) 5-azacytidine (AZ) or dimethyl sulfoxide (DMSO); (2) a combination of growth factors involved in early cardiomyogenesis (BMP-2 + bFGF + IGF-1); (3) the Wnt signaling activators lithium chloride (LiCl) and phorbol-12-myristate-13-acetate (PMA); and (4) direct contact with neonatal rat cardiomyocytes. Expression of cardiomyocyte-specific proteins and β-catenin were assessed by quantitative RT-PCR, immunofluorescence and Western blot. Cocultures of human UCBMSCs with neonatal rat cardiomyocytes were also analyzed for the presence of calcium oscillations and changes in electrical potential using Fura Red and di-4-ANEPPS confocal imaging, respectively. Induction of cardiac-specific proteins was not detected in 5-AZ- or DMSO-treated cells. Following DMSO addition, β-catenin cytoplasmic expression increased, but did not translocate into cell nuclei to promote cardiac gene activation. Likewise, neither co-stimulation with BMP-2 + bFGF + IGF-1, nor exposure to LiCl and PMA resulted in the acquisition of a cardiac phenotype by UCBMSCs. Direct contact with neonatal rat cardiomyocytes promoted neither the expression of cardiomyocyte-specific proteins, nor the presence of calcium rhythmic oscillations and potential-dependent fluorescence emission in UCBMSCs. The cardiomyogenic stimuli investigated in this study failed to transdifferentiate human UCBMSCs. Alternative strategies or regulatory factors and signaling pathways may be better suited to recruit UCBMSCs into cardiac cell lineage.

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Acknowledgments

The authors specially thank Anna Llach and Montse Barriga for confocal imaging assistance. This work was supported by Ministerio de Educación y Ciencia (SAF 2004-08044-C03-01 and SAF2008-05144-C02-01) and Fundació Marató TV3 (2007, Malalties Cardiovasculars). We also appreciate support from Fundació Daniel Bravo Andreu and Fundació Roviralta. The authors thank the reviewers and editors for invaluable advice.

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Authors and Affiliations

  1. ICREC (Heart Failure and Cardiac Regeneration) Program, Cardiology Service, Hospital de la Santa Creu i Sant Pau, ICCC, C/Sant Antoni Mª Claret 167, 08025, Barcelona, Spain

    Santiago Roura, Cristina Prat-Vidal, Carolina Soler-Botija, Carolina Gálvez-Montón & Antoni Bayes-Genis

  2. Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Barcelona, Spain

    Jordi Farré & Marta Vilalta

  3. Cardiovascular Research Center (CSIC-ICCC), Barcelona, Spain

    Jordi Farré, Leif Hove-Madsen & Marta Vilalta

  4. Department of Medicine, UAB, Barcelona, Spain

    Antoni Bayes-Genis

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  1. Santiago Roura
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395_2009_81_MOESM1_ESM.tif

Supplementary Fig. 1 Coculture of human ATDPCs with neonatal rat cardiomyocytes: acquisition of cardiac-specific proteins. Representative merge images showing the intensity and disposition of sarcomeric α-actinin, β-MHC, Cx43, GATA-4, cTnI and Nkx2.5, which were comparable to those of the surrounding neonatal cardiomyocytes, in human GFP+ ADTPCs (green). The cytoplasmic disposition of cTnI within human GFP+ cells evoked the subcellular sarcomeric organization observed in neonatal cardiomyocytes in culture. N=40 microscopic fields per condition were analysed in two independent experiments. Scale bars: 20 μm (TIFF 3560 kb)

395_2009_81_MOESM2_ESM.tif

Supplementary Fig. 2 Coculture of human UCBMSCs with neonatal rat cardiomyocytes: distribution of Cx-43. Representative merge images from independent experiments (n=2), in which at least 20 microscopic fields were studied, showing the distribution of Cx-43 between human GFP+ UCBMSCs (green) and surrounding rat cardiomyocytes. Cx-43 displays an heterogeneous and sparse location in UCBMSC border compared to adjacent cardiomyocytes (white arrowheads). Inset in the right upper corner details distribution of Cx-43 in GFP+ UCBMSC-to-cardiomyocyte contacts. Scale bars: 20 μm (TIFF 3449 kb)

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Roura, S., Farré, J., Hove-Madsen, L. et al. Exposure to cardiomyogenic stimuli fails to transdifferentiate human umbilical cord blood-derived mesenchymal stem cells. Basic Res Cardiol 105, 419–430 (2010). https://doi.org/10.1007/s00395-009-0081-8

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