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Conversion of Human Induced Pluripotent Stem Cells into Cardiac Progenitor Cells for Heart Repair
Author(s) -
Xuan Wanling,
Ali Ailia,
Ashraf Muhammad
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.978.3
Subject(s) - induced pluripotent stem cell , microbiology and biotechnology , immunofluorescence , mef2c , biology , progenitor cell , stem cell , myocyte , cell culture , transplantation , chemistry , immunology , antibody , embryonic stem cell , transcription factor , gene , medicine , biochemistry , genetics
Objective Cardiac progenitor cells (CPCs) may offer a promising avenue for cardiac repair due to their ability to proliferate and multiply. However, generating abundant and functional autologous CPCs for transplantation remains to be a significant challenge. Here, we explore a novel strategy for human CPCs generation from human induced pluripotent stem cells (iPSCs) in large numbers for their high reproducibility, high purity, and ease of production. Methods and Results We screened for small molecules, which highly induced Nkx2.5 expression in Human iPSC cell line (ACS‐1021™) by RT‐PCR. A novel small molecule MD1 was identified, that dramatically induced the expression of CPCs related genes (Nkx2.5, GATA4, Tbx5, ISL‐1 and Mef2c) for 7‐day treatment in monolayer human iPSC. Briefly, monolayer iPSCs were cultured in RPMI/B27 minus insulin medium supplemented with MD1(20μM) for 7 days. Immunofluorescence staining demonstrated that cardiac transcription factors (Nkx2.5, GATA4 and ISL‐1) were highly expressed in hiPSC with MD1 treatment. FACS analysis showed that about 96.5±2.5% cells were Nkx2.5 positive in MD1 treated human iPSC. Furthermore, these Nkx2.5 positive cells were multipotent and directly differentiated into all three cardiovascular lineages, including cardiomyocytes (CMs), endothelia cells (ECs) and smooth muscle cells (SMCs) in basal differentiation conditions without any specific induction signaling molecules. These cardiac cells expressed CM‐, EC‐ and SMC‐specific proteins detected by immunofluorescence staining. FACS analysis revealed a high proportion of cardiac lineage cells from these induced CPC. We were able to obtain about 95.2±2.1% CMs, 90.3±2.5% ECs and 92.3±1.8% SMCs in basal differentiation medium respectively. In addition, we demonstrated the differentiated ECs exhibited a similar phenotype and function like primary ECs. Moreover, these induced CPCs exhibited cytoprotective effects under hypoxic condition. Conclusion We successfully generated highly pure human CPCs in large numbers from iPSC by a single small molecule, which might be an efficient methodology for providing an unlimited source of cells for cardiac regeneration. Support or Funding Information NIH grant HL126516