Clonal Differentiation of Skeletal Muscle–Derived CD34−/45− Stem Cells Into Cardiomyocytes In Vivo
| Autor: | Tetsuro Tamaki, Masahiro Nitta, Yoshiyasu Uchiyama, Maki Masuda, Akio Hoshi, Akira Akatsuka, Yoshinori Okada, Kayoko Tono |
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| Rok vydání: | 2010 |
| Předmět: |
Cellular differentiation
LIM-Homeodomain Proteins Muscle Fibers Skeletal Myocardial Infarction Gene Expression Antigens CD34 Mice Transgenic Biology Mice Cellular cardiomyoplasty Basic Helix-Loop-Helix Transcription Factors medicine Animals Myocyte Myocytes Cardiac Cells Cultured Homeodomain Proteins Myocardium Stem Cells Cardiac muscle Gap Junctions Skeletal muscle Cell Differentiation Cell Biology Hematology Cell sorting Molecular biology Actins GATA4 Transcription Factor medicine.anatomical_structure Myogenic Regulatory Factors Cell culture Homeobox Protein Nkx-2.5 Leukocyte Common Antigens Stem cell Biomarkers Stem Cell Transplantation Transcription Factors Developmental Biology |
| Zdroj: | Stem Cells and Development. 19:503-512 |
| ISSN: | 1557-8534 1547-3287 |
| DOI: | 10.1089/scd.2009.0179 |
| Popis: | The differentiation and/or therapeutic potential of skeletal muscle-derived stem cells for cardiac infarction have been studied extensively for use in cellular cardiomyoplasty, as injured cardiomyocytes exhibit limited regenerative capacity. We previously reported cardio-myogenic differentiation of skeletal muscle-derived CD34+/45(-) (Sk-34) stem cells after therapeutic transplantation. However, the clonal differentiation potential of these cells remains unknown. Here, we show that skeletal muscle-derived CD34(-)/45(-) (Sk-DN) stem cells, which are situated upstream of Sk-34 cells in the same lineage, exhibit clonal differentiation into cardiomyocytes after single cell-derived single-sphere implantation into myocardium. Sk-DN cells were enzymatically isolated from green fluorescent protein (GFP) transgenic mice and purified by flow cytometry, and were then clonally cultured in collagen-based medium with bFGF and EGF after clonal cell sorting. Single cell-derived single-sphere colonies of Sk-DN cells were directly implanted into the wild-type mouse myocardium. At 4 weeks after implantation, donor cells exhibited typical cardiomyocyte structure with the formation of gap-junctions between donor and recipient cells. Expression of specific mRNAs for cardiomyocytes, such as cardiac actin and GATA-4, Nkx2-5, Isl-1, Mef2, and Hand2, were also seen in clonal cell cultures of Sk-DN cells. Cell fusion-independent differentiation was also confirmed by bulk cell transplantation using Cre- and loxP (enhanced GFP)-mice. We conclude that Sk-DN cells can give rise to cardiac muscle cells clonally, and that skeletal muscle includes a practical cell source for cellular cardiomyoplasty. |
| Databáze: | OpenAIRE |
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