Cell-Surface Marker Signature for Enrichment of Ventricular Cardiomyocytes Derived from Human Embryonic Stem Cells
| Autor: | Nil Emre, Jeffrey M. Hegarty, Kirk U. Knowlton, Alec D. Witty, Canzhao Liu, Dekker C. Deacon, Kelly A. Frazer, Jason G. Vidal, Elie Farah, Karina Palomares, Sylvia M. Evans, Mirko Corselli, Christian T. Carson, Patrick van Vliet, Maggie Zhu, Ralf J. Dirschinger, Eric Adler, Jennifer Veevers, Kunfu Ouyang, Jonathan D. Grinstein, Neil C. Chi, Ju Chen, Jody Martin |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Human Embryonic Stem Cells Cell cardiac differentiation Regenerative Medicine Cardiovascular Biochemistry Green fluorescent protein Myocytes Cardiac cell-surface marker signature Induced pluripotent stem cell lcsh:QH301-705.5 Cells Cultured lcsh:R5-920 Cultured Trihexosylceramides Cell Differentiation CD Cell biology Heart Disease medicine.anatomical_structure Development of treatments and therapeutic interventions lcsh:Medicine (General) Cardiac Biotechnology Myosin Light Chains Myosin light-chain kinase Heart Ventricles Cells 1.1 Normal biological development and functioning Transgene Clinical Sciences Biology Cell Line 03 medical and health sciences ventricular cardiomyocytes Antigens CD Underpinning research Genetics medicine Humans Antigens Stem Cell Research - Embryonic - Human Myocytes Bacterial artificial chromosome 5.2 Cellular and gene therapies Cell Biology Stem Cell Research Embryonic stem cell 030104 developmental biology MYL2 lcsh:Biology (General) Biochemistry and Cell Biology Cardiac Myosins Developmental Biology |
| Zdroj: | Stem cell reports, vol 11, iss 3 Stem Cell Reports, Vol 11, Iss 3, Pp 828-841 (2018) |
| ISSN: | 2213-6711 |
| Popis: | Summary: To facilitate understanding of human cardiomyocyte (CM) subtype specification, and the study of ventricular CM biology in particular, we developed a broadly applicable strategy for enrichment of ventricular cardiomyocytes (VCMs) derived from human embryonic stem cells (hESCs). A bacterial artificial chromosome transgenic H9 hESC line in which GFP expression was driven by the human ventricular-specific myosin light chain 2 (MYL2) promoter was generated, and screened to identify cell-surface markers specific for MYL2-GFP-expressing VCMs. A CD77+/CD200− cell-surface signature facilitated isolation of >97% cardiac troponin I-positive cells from H9 hESC differentiation cultures, with 65% expressing MYL2-GFP. This study provides a tool for VCM enrichment when using some, but not all, human pluripotent stem cell lines. Tools generated in this study can be utilized toward understanding CM subtype specification, and enriching for VCMs for therapeutic applications. : In this article, Evans and colleagues generated an H9 BAC transgenic reporter cell line and performed a flow cytometry screen to identify a cell-surface signature specific for MYL2-GFP-expressing VCMs. The cell-surface signature, CD77+/CD200−, facilitated isolation of a nearly pure hESC-derived CM population, enriched for VCMs. VCM enrichment was achieved when using some, but not all, human pluripotent stem cell lines. Tools generated in this study serve to advance our understanding of CM subtype specification, commitment, and maturation. Keywords: cardiac differentiation, human embryonic stem cells, ventricular cardiomyocytes, cell-surface marker signature |
| Databáze: | OpenAIRE |
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