Cloning of human cord blood-mesenchymal stem cells for isolation of enriched cell population of higher proliferation and differentiation potential
| Autor: | Faten S. Mahmoud, Shereen Shawky, Zeinab Demerdash, Marwa Hassan, Rania H. Khalifa, Noha Ali, Salwa H. Mohamed, Hanan El Baz |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Homeobox protein NANOG Population Biology Mesenchymal Stem Cell Transplantation Immunophenotyping Umbilical Cord 03 medical and health sciences Kruppel-Like Factor 4 0302 clinical medicine SOX2 Genetics Humans Progenitor cell Cloning Molecular education Molecular Biology Cells Cultured Cell Proliferation education.field_of_study Adipogenesis Mesenchymal stem cell Reproducibility of Results Cell Differentiation Mesenchymal Stem Cells General Medicine Fetal Blood Cell biology 030104 developmental biology KLF4 030220 oncology & carcinogenesis Cord blood Chondrogenesis |
| Zdroj: | Molecular biology reports. 47(5) |
| ISSN: | 1573-4978 |
| Popis: | Heterogeneity of Mesenchymal stem cells (MSCs) imposes limitations for their in vitro expansion and accounts for the lack of reproducibility in some clinical studies. So, this study was designed to isolate and enrich clones of multipotent and self-renewing MSCs from cord blood (CB). Enriched clones with higher proliferation and differentiation potential provide regenerative cells suitable for various clinical demands. MSCA and MSCB original (progenitor) cells were isolated from CB samples, and single cells were cloned by limiting dilution method, in mouse embryonic fibroblast conditioned media. Original MSCs and their single-cell derived clones were characterized by identifying their proliferation rate, immunophenotyping of surface antigens, expression of pluripotency and proliferation genes (Oct4, Sox2, Nanog, KLF4, c-Myc, and PDGFRA), and differentiation potential into multiple lineages (osteogenic, adipogenic, and chondrogenic). Some single-cell clones of MSCA showed a higher proliferation rate and greater differentiation potential than their original cells. However, original MSCB cells were of greater proliferation and differentiation potential than their derived single-cell clones, except for one clone which had comparable results. Cloning of MSCs was attainable when cultured in mouse embryonic fibroblast conditioned media. Single clones with higher proliferation and differentiation potential than their original progenitor cells were obtained by cloning of poorly functioning MSCs progenitor cells, enabling the selection of more therapeutically efficacious MSCs with better performance in clinical applications. Moreover, this study draws attention to the importance of CD105 as a possible MSCs biomarker associated with the multilineage commitment of MSCs. |
| Databáze: | OpenAIRE |
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