Induction of cardiac myogenic lineage development differs between mesenchymal and satellite cells and is accelerated by bone morphogenetic protein-4

Abstract

Our aim was to further elucidate the cardiac lineage development of bone marrow-derived mesenchymal stem cells (MSC) and to identify cells which had the potential for cardiac myogenic differentiation when compared to skeletal muscle satellite (Sk-sat) myogenesis. Unlike Sk-sat, MSC expressed the early cardiac markers Nkx2.5 and GATA4. Their expression was significantly increased by culturing MSC with Bone Morphogenetic Protein 4 (BMP4). Enhanced cardiac myogenic lineage differentiation and loss of stem cell characteristics induced by BMP4 were further confirmed by flow cytometry of cells stained for Nkx2.5 and Sca-1 expression. MSC also expressed skeletal genes (MyoG, ssTnI, Sk-Act) early in culture but their expression was suppressed when BMP4 was added from day 0–6 (p < 0.05). BMP4 treated MSC also exhibited a 6-fold increase in cTnI expression by day 12 in culture. The average MSC action potential time duration at 90% (APD90) was 32.3 ± 4 ms, with some cells exhibiting action potentials closer to Sk-sat APD90 of 13.7 ± 0.9 ms. After treatment with BMP4, MSC significantly increased their APD90 to 54.4 ± 7.6 ms, shifting from the shorter skeletal-like signature, towards a longer action potential duration more characteristic of a cardiomyocyte signature. Our results show that MSC and Sk-sat exhibit similarities in myogenic lineage development early in culture but that BMP4 clearly enhances cardiac myogenic development, suppresses skeletal myogenesis, and leads to loss of “stemness” in MSC. These findings provide novel information regarding the use of BMP4 to accelerate cardiac myogenic development in harvested MSC and further support the use of MSC in cardiac regenerative therapy.

BMP4 inhibits myogenic differentiation of bone marrow–derived mesenchymal stromal cells in mdx mice

“…This suggested that BMP4 has an inhibitory role in the myogenic differentiation of BMSCs after 5-Aza treatment. This was consistent with previous reports by Grajales et al [15] and Kennedy et al [16], who reported that BMP4 suppressed skeletal myogenic differentiation. The BMP4 intracellular signaling pathway involves a specific set of receptor-mediated Smad proteins, and the induction of Smads was shown to negatively regulate myogenic differentiation of C2C12 myoblasts, embryonic stem cells and mesenchymal stromal cells [12].…”

“…More recently, it was revealed that BMP4 also modulates myogenic differentiation in BMSCs. Grajales et al [15] reported that BMP4 enhanced cardiac myogenic development and suppressed skeletal myogenesis, leading to loss of "stemness" in BMSCs. Another study showed that retinoic acid enhanced skeletal myogenesis in stem cells by inhibition of BMP signaling [16].…”

Commitment of Satellite Cells Expressing the Calcium Channel α2δ1 Subunit to the Muscle Lineage

“…The number of independent cell cultures analyzed was a minimum of 3, and the same gene/cDNA mix was analyzed 3 times in qPCR 96 well plates. All genes were referenced to the geometric mean of at least two control genes selected from YWHAZ, 18S, and HPRT1 [12, 13]. The primer sequences are given in Table 1.…”

Cellular Cardiomyoplasty Using Skeletal Muscle Stem Cells

“…Using inbred animals in cellular cardiomyoplasty, it is possible to use the male donor cells transplanted into female animals and identify the donor cells by the presence of Y chromosome. The differentiation fate of the transplanted cells can be determined by the specific cell markers by histologic or immunohistochemical procedures [60]…”

“…Alternatively induction of cardiac fibroblasts into cardiomyocyte-like cells [134][135][136][137][138][139] can be another way of myocardial regeneration. Cardiomyogenic pretreatment significantly increased the formation of cardiac myocytes after their transplantation into the injured heart [140] and different stem cells may require different treatment [141,142].…”