Rho-associated protein kinases play an important role in the differentiation of rat adipose-derived stromal cells into cardiomyocytes in vitro

• Published in: PloS one Vol. 9; no. 12; p. e115191
• Main Authors: Zhao, Lili, Yang, Gongshe, Zhao, Xin
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 18.12.2014; Public Library of Science (PLoS)
• Subjects: 1-Phosphatidylinositol 3-kinase; Adipose Tissue - cytology; AKT protein; Animal sciences; Animals; Biology and life sciences; c-Jun protein; Calcium; Calcium Signaling; Cardiomyocytes; Cell Differentiation; Cells, Cultured; Cytoskeleton; Cytoskeleton - metabolism; Differentiation; Disease; Growth factors; Heart attacks; Heart cells; Heart diseases; Heart failure; JNK Mitogen-Activated Protein Kinases - metabolism; JNK protein; Kinases; Male; Medicine and Health Sciences; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mesenchyme; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - physiology; Phosphorylation; Physiology; Protein Kinase Inhibitors - pharmacology; Protein kinases; Proto-Oncogene Proteins c-akt - metabolism; Rats; Rats, Sprague-Dawley; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - genetics; rho-Associated Kinases - metabolism; Rocks; Rodents; Signaling; siRNA; Stem cell transplantation; Stem cells; Stromal cells; Transcription factors; Transplantation; Zoology; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0115191

Adipose-derived stromal cells (ADSCs) represent a readily available abundant supply of mesenchymal stem cells and have the ability to differentiate into cardiomyocytes in mice and human, making ADSCs a promising source of cardiomyocytes for transplantation. However, there has been no report of differentiation of rat ADSCs into cardiomyocytes. In addition, signaling pathways in the differentiation process from ADSCs to cardiomyocytes are unknown. In this study, we first demonstrated that rat ADSCs spontaneously differentiated into cardiomyocytes in vitro, when cultured on a complete medium formulation MethoCult GF M3534. These differentiated cells possessed cardiomyocyte phenotype and expressed cardiac markers. Moreover, these cells showed open excitation-contracting coupling and Ca2+ transient and contracted spontaneously. The role of Rho-associated protein kinases (ROCKs) in the differentiation process was then studied by using ROCK-specific inhibitor Y-27632 and ROCK siRNAs. These agents changed the arrangement of cytoskeleton and diminished appearance of cardiomyocyte phenotype, accompanied by inhibition of c-Jun N-terminal kinase (JNK) phosphorylation and promotion of Akt phosphorylation. Collectively, this is the first study to demonstrate that rat ADSCs could spontaneously differentiate into cardiomyocytes in vitro and ROCKs play an important role in the differentiation of ADSCs into beating cardiomyocytes in conjunction of the PI3K/Akt pathway and the JNK pathway.