Human Embryonic and Fetal Mesenchymal Stem Cells Differentiate toward Three Different Cardiac Lineages in Contrast to Their Adult Counterparts

Abstract

Mesenchymal stem cells (MSCs) show unexplained differences in differentiation potential. In this study, differentiation of human (h) MSCs derived from embryonic, fetal and adult sources toward cardiomyocytes, endothelial and smooth muscle cells was investigated. Labeled hMSCs derived from embryonic stem cells (hESC-MSCs), fetal umbilical cord, bone marrow, amniotic membrane and adult bone marrow and adipose tissue were co-cultured with neonatal rat cardiomyocytes (nrCMCs) or cardiac fibroblasts (nrCFBs) for 10 days, and also cultured under angiogenic conditions. Cardiomyogenesis was assessed by human-specific immunocytological analysis, whole-cell current-clamp recordings, human-specific qRT-PCR and optical mapping. After co-culture with nrCMCs, significantly more hESC-MSCs than fetal hMSCs stained positive for α-actinin, whereas adult hMSCs stained negative. Furthermore, functional cardiomyogenic differentiation, based on action potential recordings, was shown to occur, but not in adult hMSCs. Of all sources, hESC-MSCs expressed most cardiac-specific genes. hESC-MSCs and fetal hMSCs contained significantly higher basal levels of connexin43 than adult hMSCs and co-culture with nrCMCs increased expression. After co-culture with nrCFBs, hESC-MSCs and fetal hMSCs did not express α-actinin and connexin43 expression was decreased. Conduction velocity (CV) in co-cultures of nrCMCs and hESC-MSCs was significantly higher than in co-cultures with fetal or adult hMSCs. In angiogenesis bioassays, only hESC-MSCs and fetal hMSCs were able to form capillary-like structures, which stained for smooth muscle and endothelial cell markers.Human embryonic and fetal MSCs differentiate toward three different cardiac lineages, in contrast to adult MSCs. Cardiomyogenesis is determined by stimuli from the cellular microenvironment, where connexin43 may play an important role.

Brief Report: The Potential Role of Epigenetics on Multipotent Cell Differentiation Capacity of Mesenchymal Stromal Cells

“…These data are in agreement with previous studies demonstrating enhanced trilineage differentiation capacity of HUCPVCs [4,16]. Human MSCs derived from embryonic and fetal sources have also exhibited the ability to enhance cardiomyogenesis compared with their adult counterparts [17]. Moreover, several reports have recently demonstrated the regenerative potential of HUCPVCs in different animal models [18][19][20].…”

Gap Junctional Coupling with Cardiomyocytes is Necessary but Not Sufficient for Cardiomyogenic Differentiation of Cocultured Human Mesenchymal Stem Cells

“…Cx43;(1) fetal AM hMSCs transduced with an LV-encoding eGFP were used as control cells (Cx43;(1) þ eGFP fetal AM hMSCs) in these experiments. For a detailed description of Materials and Methods, the reader is referred to Supporting Information and previous papers of our research group [19][20][21].…”

“…However, to what extent MSCs can undergo cardiac differentiation and which factors are involved in this process are still unclear [16][17][18]. Although donor age and heterocellular interactions were found to affect the cardiomyogenic differentiation potential of MSCs [19], more research is needed into their biological properties and the modification thereof to improve the therapeutic potential of MSC-based cardiac cell therapy.…”

“…Accordingly, in this study, we specifically investigated the role of Cx43, the major gap junction protein of the working myocardium, in cardiomyogenic differentiation of naturally Cx43-rich fetal and Cx43-poor adult human MSCs (hMSCs) [19] in cocultures with neonatal rat ventricular CMCs (nrCMCs). To this end, Cx43 expression levels in hMSCs were either downregulated by short hairpin RNA (shRNA)-mediated RNA interference (RNAi) or upregulated by recombinant human Cx43 (hCx43) gene delivery.…”

Functional properties of a Brazilian derived mouse embryonic stem cell line

“…The cell-cell coupling between ESC and cardiomyocytes have been observed by several groups using the model of co-culture seeding rats cardiomyocytes and different sources of stem cells (Ramkisoensing et al 2011, Bollini et al 2011, Valarmathi et al 2011). Here we used the co-culture with mouse eGPF-cardiomyocytes and USP-1 mESC to test the hypothesis whether both types of cells are able to make functional GJIC.…”