Biopolym. Cell. 2014; 30(6):469-476.
Molecular and Cell Biotechnologies
Biological properties of neural crest-derived
multipotent stem cells from the bulge region of whisker follicle expanded in new culture conditions
- Institute of Genetic and Regenerative Medicine, NAMS of Ukraine
67, Vyshgorodska Str., Kyiv, Ukraine, 04114 - Biotechnology laboratory ilaya.regeneration, Medical company ilaya
9, I. Kramskogo Str., Kyiv, Ukraine, 03115 - Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
Abstract
Aim. The work is aimed at obtaining the culture of neural crest-derived multipotent stem cells (NC-MSCs) in new culture conditions and to investigate their biological properties. Methods. NC-MSCs were grown from the explants of the bulge region of whisker follicle of adult mice. The cell cultures were examined by the following methods: sphere-forming assay, directed multilineage differentiation, CFU assay, immunocytochemistry, flow cytometry, RT-PCR. Results. The obtained NC-MSCs expressed the typical neural crest markers (nestin, Sox10 and Sox2) and were differentiated into adipocytes, osteoblasts and Schwann cells. Under our original growing conditions, the culture of NC-MSCs at the third passage had the following parameters: 66.8 % nestin+, 3.1 % ALDH brigth and 33.3 % clonogenic cells. The NC-MSCs growth rate depended on plating density. EGF and bFGF demonstrated a dose-dependent mitogenic action on NC-MSCs. Conclusions. The proposed approach permits the NC-MSC expansion with the maintenance of their main functional properties. Further optimization of the culture conditions will be based on the use of growth factors and low plating density.
Keywords: neural crest, multipotent stem cells, colony-forming units, plating density, growth factors
Full text: (PDF, in English)
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