Dynamic mitochondrial changes during differentiation of P19 embryonic carcinoma cells into cardiomyocytes

Abstract

Murine P19 embryonal carcinoma cells are multipotent cells that can differentiate into cardiomyocytes when treated with dimethyl sulfoxide. This experimental model provides an invaluable tool to study different aspects of cardiac differentiation, such as the function of cardiac‑specific transcription factors and signaling pathways, and the regulation of contractile protein expression. The role of mitochondria during cardiac differentiation is unclear. In this context, we have examined the mitochondrial-related changes in undifferentiated and differentiated P19 cells. We observed that mitochondrial DNA content sharply decreased in P19 cell aggregates compared to undifferentiated cells, accompanied by decreased levels of adenosine triphosphate (ATP) and reactive oxygen species (ROS). Following the aggregation stage, the mitochondrial DNA content reached its highest level on day 7 of the differentiation process, with the intracellular ROS level showing a trend to increase, similar to cellular ATP production. In conclusion, our study on differentiating P19 embryonal carcinoma cells provides new insights into the role of mitochondria in the differentiation of P19 stem cells into beating cardiomyocytes.

The Calcineurin-Drp1-Mediated Mitochondrial Fragmentation Is Aligned with the Differentiation of c-Kit Cardiac Progenitor Cells

“…Our results are consistent with the findings of other studies whereby the use of pharmacological nitric oxide (NO) donors to increase mitochondrial numbers promotes maturation of fastdividing mesoangioblasts into cardiomyocytes whilst decreasing mitochondrial content using respiratory chain inhibitors and chloramphenicol perturbs cardiomyocyte differentiation in slow-dividing populations (21). Rounding of the mitochondria was also observed in the aggregation stage of P19 cells differentiation into cardiomyocytes (22). Knockdown of Drp1 negatively influences 30-days neurogenesis of ESCs, coincident with a delayed reduction of Oct4 and Nanog during the mid-differentiation process (23).…”

Gene-Specific Assessment of Guanine Oxidation as an Epigenetic Modulator for Cardiac Specification of Mouse Embryonic Stem Cells

“…For example, low levels of H 2 O 2 stimulated cardiomyogenesis of ESCs and induced proliferations of cardiomyocytes derived from ESCs as investigated by nuclear translocation of cyclin D1, down-regulation of p27 (Kip1), and retinoblastoma phosphorylation, and increased of Ki-67 expression [ 46 ]. Furthermore, intracellular ROS shows a trend to increase during the process of ESC differentiation and has an essential role in cardiomyocyte differentiation of ESCs [ 28 , 47 – 49 ]. These findings imply that ROS can act as signals for cardiomyocyte differentiation and growth.…”

Identification of differential microRNAs and messenger RNAs resulting from ASXL transcriptional regulator 3 knockdown during during heart development

“…CRL-1825) and cultured at 37°C under 5% CO 2 in alpha-minimum essential medium (α-MEM; Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (Gibco, Waltham, MA, USA). As described in previous studies [ 32 ], P19 cells were stimulated with 1% DMSO (Sigma, D4540) for 4 days to induce differentiation. Then, 30–40 embryo-like bodies were inoculated into a six-well plate with α-MEM for further culture.…”