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
1, 2Vasyliev R. G., 1, 2Rodnichenko A. E., 1, 2Zubov D. A., 3Rymar S. Y., 3Gubar O. S., 1Labunets I. F., 1Novikova S. N.
  1. Institute of Genetic and Regenerative Medicine, NAMS of Ukraine
    67, Vyshgorodska Str., Kyiv, Ukraine, 04114
  2. Biotechnology laboratory ilaya.regeneration, Medical company ilaya
    9, I. Kramskogo Str., Kyiv, Ukraine, 03115
  3. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


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


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