Autor: |
Samoilova EM; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia. samoyket@gmail.com., Revkova VA; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia., Brovkina OI; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia., Kalsin VA; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia., Melnikov PA; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia.; Department of Fundamental and Applied Neurobiology, V. P. Serbsky Federal Medical Research Center for Psychiatry and Narcology, Ministry of Health of the Russian Federation, Moscow, Russia., Konoplyannikov MA; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia.; Institute of Regenerative Medicine, I. M. Sechenov First Moscow State Medical University (Sechenov University), Moscow, Russia., Galimov KR; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia., Nikitin AG; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia., Troitskiy AV; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia., Baklaushev VP; Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of Russia, Moscow, Russia. |
Abstrakt: |
In in vitro experiments on cultures of human multipotent stem cells from the human bonearrow and dental pulp, we studied direct reprogramming towards neuro-glial lineage cells using a cocktail of small molecules. Reprogramming by the previously published protocol (with a cocktail containing β-mercaptoethanol, LIF, VPA, CHIR99021, and RepSox) and by the optimized protocol (VPA, RG108, А83-01, dorsomorphin, thiazovivin, CHIR99021, forskolin, and Isx9) allows obtaining cells with immunophenotypic and genetic signs of neural stem cells. However, neither the former, nor the optimized protocols allowed preparing neural progenitors capable of adequate terminal differentiation from both bone marrow-derived mesenchymal stem cells and nestin-positive neural crest-derived mesenchymal stem cells. Real-time PCR demonstrated the expression of some neurogenesis markers, but neural stem cell-specific expression pattern was not observed. The findings lead us to a conclusion that reprogramming with small molecules without additional factors modifying gene expression does not allow reproducible production of human neural stem cell-like progenitors that can be used as the source of neural tissue for the regenerative therapy. |