PMID- 30196409 OWN - NLM STAT- MEDLINE DCOM- 20190423 LR - 20190423 IS - 1940-6029 (Electronic) IS - 1064-3745 (Print) IS - 1064-3745 (Linking) VI - 1842 DP - 2018 TI - Isolation, Characterization and Differentiation of Mouse Cardiac Progenitor Cells. PG - 183-191 LID - 10.1007/978-1-4939-8697-2_12 [doi] AB - Despite several strategies developed for replenishing the dead myocardium after myocardial infarction (MI), stem cell therapy remains the leading method to regenerate new myocardium. Although induced pluripotent stem cells (iPS) and transdifferentiation of the differentiated cells have been used as novel approaches for myocardial regeneration, these approaches did not yield very successful results for myocardial regeneration in in vivo studies. Asynchronous contractility of newly formed cardiomyocytes with the existing cardiomyocytes is the most important issue with iPS approach, while very low yield of transdifferentiated cardiomyocytes and their less chances to beat in the same rhythm as existing cardiomyocytes in the MI heart are important caveats with transdifferentiation approach. CSCs are present in the heart and they have the potential to differentiate into myocardial cells. However, the number of resident CSCs is very low. Therefore, it is important to get maximum yield of CSCs during isolation process from the heart. Increasing the number of CSCs and initiating their differentiation ex vivo are crucial for CSC-based stem cell therapy. Here, we present a better method for isolation, characterization and differentiation of CSCs from the mouse heart. We also demonstrated morphological changes in the CSCs after 2 days, 3 days, and 7 days in maintenance medium and a separate group of CSCs cultured for 12 days in differentiation medium using Phase-Contrast microscopy. We have used different markers for identification of CSCs isolated from the mouse heart such as marker for mouse CSC, Sca-1, cardiac-specific markers NKX2-5, MEF2C, GATA4, and stemness markers OCT4 and SOX2. To characterize the differentiated CSCs, we used CSCs maintained in differentiation medium for 12 days. To evaluate differentiation of CSCs, we determined the expression of cardiomyocyte-specific markers actinin and troponin I. Overall; we described an elegant method for isolation, identification, differentiation and characterization of CSCs from the mouse heart. FAU - Yadav, Santosh Kumar AU - Yadav SK AD - Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, NE, USA. FAU - Mishra, Paras Kumar AU - Mishra PK AD - Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, NE, USA. paraskumar.mishra@unmc.edu. AD - Department of Anesthesiology, University of Nebraska Medical Center, Omaha, NE, USA. paraskumar.mishra@unmc.edu. LA - eng GR - R01 HL113281/HL/NHLBI NIH HHS/United States GR - R01 HL116205/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PL - United States TA - Methods Mol Biol JT - Methods in molecular biology (Clifton, N.J.) JID - 9214969 RN - 0 (Biomarkers) SB - IM MH - Animals MH - Biomarkers MH - *Cell Differentiation MH - *Cell Separation MH - Fluorescent Antibody Technique MH - Immunophenotyping/methods MH - Mice MH - Myoblasts, Cardiac/*cytology/*metabolism MH - Myocytes, Cardiac/cytology/metabolism MH - *Phenotype PMC - PMC6206846 MID - NIHMS993586 OTO - NOTNLM OT - Actinin OT - CSC characterization OT - CSC differentiation OT - Cardiac stem cells OT - Sca-1 OT - Troponin I EDAT- 2018/09/10 06:00 MHDA- 2019/04/24 06:00 PMCR- 2019/01/01 CRDT- 2018/09/10 06:00 PHST- 2018/09/10 06:00 [entrez] PHST- 2018/09/10 06:00 [pubmed] PHST- 2019/04/24 06:00 [medline] PHST- 2019/01/01 00:00 [pmc-release] AID - 10.1007/978-1-4939-8697-2_12 [doi] PST - ppublish SO - Methods Mol Biol. 2018;1842:183-191. doi: 10.1007/978-1-4939-8697-2_12.