BMP induction of cardiogenesis in P19 cells requires prior cell–cell interaction(s)

Abstract

Mouse P19 embryonal carcinoma cells undergo cardiogenesis in response to high density and DMSO. We have derived a clonal subline that undergoes cardiogenesis in response to high density, but without requiring exposure to DMSO. The new subline retains the capacity to differentiate into skeletal muscle and neuronal cells in response to DMSO and retinoic acid. However, upon aggregation, these Oct 4-positive cells, termed P19-SI because they "self-induce" cardiac muscle, exhibit increased mRNAs encoding the mesodermal factor Brachyury, cardiac transcription factors Nkx 2.5 and GATA 4, the transcriptional repressor Msx-1, and cytokines Wnt 3a, Noggin, and BMP 4. Exposure of aggregated P19-SI cells to BMP 4, a known inducer of cardiogenesis, accelerates cardiogenesis, as determined by rhythmic beating and myosin staining. However, cardiogenesis is severely inhibited when P19-SI cells are aggregated in the presence of BMP 4. These results demonstrate that cell-cell interaction is required before P19-SI cells can undergo a cardiogenic response to BMP 4. A concurrent increase in the expression of Msx-1 suggests one possible process underlying the inhibition of cardiogenesis. The phenotype of P19-SI cells offers an opportunity to explore new aspects of cardiac induction.

Crossveinless-2 Controls Bone Morphogenetic Protein Signaling during Early Cardiomyocyte Differentiation in P19 Cells

“…Bmp2 and Bmp4 were expressed from day 1 and day 2, respectively. Noggin, the specific BMP antagonist, was expressed from day 5, consistent with the previous findings (29). These data indicate that our in vitro cardiac differentiation model of P19 cells reproduces the natural course of differentiation as described in the literature (23,25), and this model is able to follow the Cv2 expression during the primary steps of cardiomyocyte differentiation.…”

Tbx20 Transcription Factor Is a Downstream Mediator for Bone Morphogenetic Protein-10 in Regulating Cardiac Ventricular Wall Development and Function

“…4A). P19 is a well characterized mouse embryo-derived teratocarcinoma cell line that can be differentiated into cardiomyocytes and is widely used to study BMP-mediated signaling (27,28). We found that Smad1 activation reached a plateau level at 50 ng/ml concentration for both of rhBMP10 and rhBmP2 in culture (Fig.…”

The primary cilium coordinates early cardiogenesis and hedgehog signaling in cardiomyocyte differentiation

“…At this time point, the individual clusters had a diameter of 0.2-0.6 mm and the clusters could be observed in small Journal of Cell Science 122 (17) networks beating synchronously at a frequency of about 60 rhythmic contractions per minute. Comparable structures of cardiogenic cell clusters were observed in differentiating P19.SI cells, another subclone of P19 cells, which contracted synchronically by about 2 weeks (Angello et al, 2007). The network between clusters in differentiated P19.CL6 cells later develop into a thick unified layer of cardiac muscle epithelium (data not shown).…”

“…Wnt and PDGF signaling are regulated by the cilium in a series of other cell types involved in growth control, migration and differentiation (reviewed by Christensen et al, 2008;Gerdes and Katsanis, 2008). BMP promotes the induction of cardiomyocytes from the mouse stem cell line P19.SI (Angello et al, 2007) and human embryonic stem cells (Takei et al, 2009). Based on their findings on Pkd2 -/-mouse embryos, Slough and colleagues (Slough et al, 2008) also hypothesized that primary cilia in the heart and/or vasculature can function as mechanosensors to Immunofluorescence microscopy analysis of acetylated tubulin (tb) on primary cilia in the heart of E11.5 WT and Ift88-null mice (tb, red; DAPI, blue).…”