Preparation of Adhesion Culture of Neural Stem/Progenitor Cells of the Olfactory Mucosa for the Treatment of Spinal Cord
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Cell Technologies in Biology and Medicine, No. 3, November, 2020
Preparation of Adhesion Culture of Neural Stem/Progenitor Cells of the Olfactory Mucosa for the Treatment of Spinal Cord Injuries O. V. Stepanova1,4, A. D. Voronova1, G. A. Fursa1,2, E. K. Karsuntseva1,2, M. P. Valikhov1,4, A. V. Chadin1, D. A. Vishnevskii1, I. V. Reshetov3, and V. P. Chekhonin1 Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 3, pp. 202-208, September, 2020 Original article submitted December 4, 2019 In this work, an optimal protocol was developed for obtaining adhesion culture of neural stem/ progenitor cells (NSPC) of rat olfactory mucosa. During the development of the protocol, the conditions for cell culturing on adhesion substrates fibronectin and laminin in DMEM/F-12 and neurobasal media with the same culture additives were compared. Cell proliferation was maximum during culturing on both substrates in the neurobasal medium. Using the immunofluorescence method, we found that culturing on fibronectin in the neurobasal medium ensured maximum (52.22%) content of nestin-positive cells in comparison with other culturing conditions. The highest percentage of βIII-tubulin-positive cells was detected in cultures growing on fibronectin in the neurobasal medium and in DMEM/F-12 (79.11 and 83.52%, respectively). Culturing in adhesion cultures in the neurobasal medium on fibronectin allowed obtaining cultures enriched with NSPC and neurons differentiating from them in a quantity sufficient for further transplantation. The developed protocol can be recommended for obtaining NPSC from human olfactory mucosa for the treatment of spinal cord injuries. Key Words: neural stem/progenitor cells; olfactory mucosa; olfactory epithelium; cell therapy; spinal cord injuries Cell therapy is a promising method for the treatment of a number of pathological conditions, in which surgery and drug therapy are ineffective. For instance, it is difficult to treat post-traumatic injuries of the spinal cord associated with the loss of a great number of neurons and destruction of neuronal pathways [6]. In experimental models and in clinical trials, the effect 1 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; 2M. V. Lomonosov Moscow State University Moscow, Russia; 3Universitty Clinical Hospital No. 1, I. M. Sechenov First Moscow State Medical University, Ministry of Health of the Russian Federation (Sechenov University), Moscow, Russia; 4Department of Clinical Diagnostics, National Medical Research Centre of Cardiology, Ministry of Health of the Russian Federation, Moscow, Russia. Address for correspondence: [email protected]. O. V. Stepanova
of various types of cells in spinal cord injuries was studied and it was shown that cell transplantation can reduce secondary damage, provide a substrate for axon growth, promote axon growth, and facilitate remyelination [12,15]. Transplantation of neural/stem proge
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