Ya-Ting Cheng1, Dong-Feng Yeih2, Shu-Man Liang1, Chia-Ying Chien1, Yen-Ling Yu1, Bor-Sheng Ko3, Yee-Jee Jan4, Cheng-Chin Kuo1, Li-Ying Sung5, Song-Kun Shyue6, Ming-Fong Chen3, Shaw-Fang Yet1, Kenneth K Wu7, Jun-Yang Liou8. 1. Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan. 2. Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan; Department of Internal Medicine, Cardinal Tien Hospital, New Taipei City, Taiwan; Department of Internal Medicine, School of Medicine, Fu-Jen Catholic University, New Taipei City, Taiwan. 3. Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan. 4. Department of Pathology and Laboratory Medicine, Taichung Veterans General Hospital, Taichung, Taiwan. 5. Institute of Biotechnology, National Taiwan University, Taipei, Taiwan. 6. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan. 7. Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan; Metabolomic Medicine Research Center, China Medical University, Taichung, Taiwan. 8. Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan; Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan. Electronic address: jliou@nhri.org.tw.
Abstract
BACKGROUND: Rho-associated kinase (ROCK) plays an important role in maintaining embryonic stem (ES) cell pluripotency. To determine whether ROCK is involved in ES cell differentiation into cardiac and hematopoietic lineages, we evaluated the effect of ROCK inhibitors, Y-27632 and fasudil on murine ES and induced pluripotent stem (iPS) cell differentiation. METHODS: Gene expression levels were determined by real-time PCR, Western blot analysis and immunofluorescent confocal microscopy. Cell transplantation of induced differentiated cells were assessed in vivo in a mouse model (three groups, n=8/group) of acute myocardial infarction (MI). The cell engraftment was examined by immunohistochemical staining and the outcome was analyzed by echocardiography. RESULTS: Cells were cultured in hematopoietic differentiation medium in the presence or absence of ROCK inhibitor and colony formation as well as markers of ES, hematopoietic stem cells (HSC) and cells of cardiac lineages were analyzed. ROCK inhibition resulted in a drastic change in colony morphology accompanied by loss of hematopoietic markers (GATA-1, CD41 and β-Major) and expressed markers of cardiac lineages (GATA-4, Isl-1, Tbx-5, Tbx-20, MLC-2a, MLC-2v, α-MHC, cTnI and cTnT) in murine ES and iPS cells. Fasudil-induced cardiac progenitor (Mesp-1 expressing) cells were infused into a murine MI model. They engrafted into the peri-infarct and infarct regions and preserved left ventricular function. CONCLUSIONS: These findings provide new insights into the signaling required for ES cell differentiation into hematopoietic as well as cardiac lineages and suggest that ROCK inhibitors are useful in directing iPS cell differentiation into cardiac progenitor cells for cell therapy of cardiovascular diseases.
BACKGROUND: Rho-associated kinase (ROCK) plays an important role in maintaining embryonic stem (ES) cell pluripotency. To determine whether ROCK is involved in ES cell differentiation into cardiac and hematopoietic lineages, we evaluated the effect of ROCK inhibitors, Y-27632 and fasudil on murine ES and induced pluripotent stem (iPS) cell differentiation. METHODS: Gene expression levels were determined by real-time PCR, Western blot analysis and immunofluorescent confocal microscopy. Cell transplantation of induced differentiated cells were assessed in vivo in a mouse model (three groups, n=8/group) of acute myocardial infarction (MI). The cell engraftment was examined by immunohistochemical staining and the outcome was analyzed by echocardiography. RESULTS: Cells were cultured in hematopoietic differentiation medium in the presence or absence of ROCK inhibitor and colony formation as well as markers of ES, hematopoietic stem cells (HSC) and cells of cardiac lineages were analyzed. ROCK inhibition resulted in a drastic change in colony morphology accompanied by loss of hematopoietic markers (GATA-1, CD41 and β-Major) and expressed markers of cardiac lineages (GATA-4, Isl-1, Tbx-5, Tbx-20, MLC-2a, MLC-2v, α-MHC, cTnI and cTnT) in murine ES and iPS cells. Fasudil-induced cardiac progenitor (Mesp-1 expressing) cells were infused into a murine MI model. They engrafted into the peri-infarct and infarct regions and preserved left ventricular function. CONCLUSIONS: These findings provide new insights into the signaling required for ES cell differentiation into hematopoietic as well as cardiac lineages and suggest that ROCK inhibitors are useful in directing iPS cell differentiation into cardiac progenitor cells for cell therapy of cardiovascular diseases.
Authors: James B Hu; Martin L Tomov; Jan W Buikema; Caressa Chen; Morteza Mahmoudi; Sean M Wu; Vahid Serpooshan Journal: Appl Phys Rev Date: 2018-12 Impact factor: 19.162
Authors: Sebastian V Rojas; George Kensah; Alexander Rotaermel; Hassina Baraki; Ingo Kutschka; Robert Zweigerdt; Ulrich Martin; Axel Haverich; Ina Gruh; Andreas Martens Journal: PLoS One Date: 2017-05-11 Impact factor: 3.240