Pellitteri Rosalia, Catania Maria Vincenza, Bonaccorso Carmela Maria, Ranno Eugenia, Dell'Albani Paola, Zaccheo Damiano
Institute of Neurological Sciences, National Research Council, Section of Catania, Catania, Italy.
J Neurosci Res. 2014 Dec;92(12):1757-66. doi: 10.1002/jnr.23442. Epub 2014 Jun 27.
Olfactory ensheathing cells (OECs) represent glial cells supporting neuronal turnover in the olfactory system. In vitro, OECs promote axonal growth as a source of neurotrophic growth factors; in vivo, they produce myelin, promoting remyelination of damaged axons. Consequently, OEC transplantation appears to be a promising treatment for spinal cord injury, although the functional recovery is limited. This might be ascribed to the microenvironment at the lesion site, lacking growth factors (GFs), nutrients, and oxygen. To mimic this condition, we used an in vitro approach by growing primary neonatal mouse OECs under hypoxic conditions and/or serum deprivation. In addition, we compared OECs survival/proliferation with that of primary cultures of Schwann cells (SCs) and astrocytes under the same experimental conditions. Cultures were analyzed by immunocytochemistry, and cell viability was evaluated by MTT assay. Different GFs, such as NGF, bFGF, and GDNF, and their combination were used to rescue cells from serum and/or oxygen deprivation. We show that the cell types were differently sensitive to the tested stress conditions and that OECs were the most sensitive among them. Moreover, OEC viability was rescued by bFGF under serum-deprived or hypoxic condition but not under conditions of drastic serum deprivation and hypoxia. bFGF was effective also for the other cell types, whereas the effect of the other GFs was negligible. This model suggests that administration of bFGF might be considered useful to sustain cell survival/proliferation after transplantation of OECs either alone or in combination with other glial cell types.
嗅鞘细胞（OECs）是支持嗅觉系统神经元更替的神经胶质细胞。在体外，OECs作为神经营养生长因子的来源促进轴突生长；在体内，它们产生髓磷脂，促进受损轴突的髓鞘再生。因此，尽管功能恢复有限，但OEC移植似乎是脊髓损伤的一种有前景的治疗方法。这可能归因于损伤部位的微环境，缺乏生长因子（GFs）、营养物质和氧气。为了模拟这种情况，我们采用体外方法，在缺氧条件下和/或血清剥夺条件下培养新生小鼠原代OECs。此外，我们在相同实验条件下比较了OECs与雪旺细胞（SCs）和星形胶质细胞原代培养物的存活/增殖情况。通过免疫细胞化学分析培养物，并通过MTT法评估细胞活力。使用不同的GFs，如NGF、bFGF和GDNF及其组合来挽救细胞免受血清和/或缺氧的影响。我们发现不同细胞类型对所测试的应激条件敏感性不同，其中OECs最为敏感。此外，在血清剥夺或缺氧条件下，bFGF可挽救OECs的活力，但在严重血清剥夺和缺氧条件下则不能。bFGF对其他细胞类型也有效，而其他GFs的作用可忽略不计。该模型表明，单独或与其他神经胶质细胞类型联合移植OECs后，给予bFGF可能有助于维持细胞存活/增殖。
