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.

中文翻译：

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嗅粘膜神经干/祖细胞粘附培养物治疗脊髓损伤的制备

在这项工作中，开发了一种最佳方案，用于获得大鼠嗅粘膜神经干/祖细胞 (NSPC) 的粘附培养。在该协议的制定过程中，比较了 DMEM/F-12 和具有相同培养添加剂的神经基础培养基中粘附基质纤连蛋白和层粘连蛋白的细胞培养条件。在神经基础培养基中的两种底物上培养期间细胞增殖是最大的。使用免疫荧光方法，我们发现与其他培养条件相比，在神经基础培养基中培养纤连蛋白可确保巢蛋白阳性细胞的最大含量 (52.22%)。在神经基础培养基和 DMEM/F-12 中的纤连蛋白上生长的培养物中检测到最高百分比的 βIII-微管蛋白阳性细胞（分别为 79.11% 和 83.52%）。在纤连蛋白上的神经基础培养基中的粘附培养物中培养允许获得富含 NSPC 的培养物和从它们分化出来的神经元，其数量足以用于进一步移植。可以推荐开发的协议从人类嗅觉粘膜中获取 NPSC 以治疗脊髓损伤。