Preparation of Adhesion Culture of Neural Stem/Progenitor Cells of the Olfactory Mucosa for the Treatment of Spinal Cord Injuries

• Published in: Bulletin of experimental biology and medicine Vol. 170; no. 1; pp. 158 - 163
• Main Authors: Stepanova, O. V., Voronova, A. D., Fursa, G. A., Karsuntseva, E. K., Valikhov, M. P., Chadin, A. V., Vishnevskii, D. A., Reshetov, I. V., Chekhonin, V. P.
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-11-2020; Springer; Springer Nature B.V
• Subjects: Biomedical and Life Sciences; Biomedicine; Cell Biology; Cell culture; Cell proliferation; Culture media; Fibronectin; Fibronectins; Immunofluorescence; Injuries; Internal Medicine; Laboratory Medicine; Laminin; Mucosa; Nestin; Neural stem cells; Neurophysiology; Olfactory epithelium; Pathology; Progenitor cells; Spinal cord injuries; Transplantation; Tubulin; olfactory epithelium; olfactory mucosa; spinal cord injuries; cell therapy; neural stem/progenitor cells
• ISSN: 0007-4888; 1573-8221
• DOI: 10.1007/s10517-020-05023-0

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.