脂肪间充质干细胞(ASCs)是成人多能干细胞,能够向中胚层谱系细胞以外的神经元分化。这项研究的目的是使用褪黑激素结合胶质细胞的条件培养基(CM)测试ASC神经分化。从健康捐赠者的脂肪抽吸物中分离出来,在经历神经分化程序之前,将ASC在基础生长培养基中扩增。为此,使用获自嗅鞘细胞和施旺细胞的CM。在一些样本中,加入1μM的褪黑激素。培养1天和7天后,使用免疫细胞化学和流式细胞术研究细胞以评估神经标记物的表达(Nestin,MAP2,突触素I,GFAP)在不同条件下。结果证实,成功的神经分化是通过神经胶质CM实现的,而单独添加褪黑激素并没有引起明显的变化。当褪黑激素与CM结合使用时,ASC神经分化增强,正如神经元标记表达的进一步改善所证明的那样,而神经胶质分化减弱。还观察到动态调制,测试褪黑素受体的表达。总之,我们的数据表明,褪黑素的神经分化能力可被有效利用,以获得神经元样分化的ASCs,用于潜在的治疗策略.
Adipose-derived mesenchymal stem cells (ASCs) are adult multipotent stem cells, able to differentiate toward neural elements other than cells of mesodermal lineage. The aim of this research was to test ASC neural differentiation using melatonin combined with conditioned media (CM) from glial cells. Isolated from the lipoaspirate of healthy donors, ASCs were expanded in a basal growth medium before undergoing neural differentiation procedures. For this purpose, CM obtained from olfactory ensheathing cells and from Schwann cells were used. In some samples, 1 µM of melatonin was added. After 1 and 7 days of culture, cells were studied using immunocytochemistry and flow cytometry to evaluate neural marker expression (Nestin, MAP2, Synapsin I, GFAP) under different conditions. The results confirmed that a successful neural differentiation was achieved by glial CM, whereas the addition of melatonin alone did not induce appreciable changes. When melatonin was combined with CM, ASC neural differentiation was enhanced, as demonstrated by a further improvement of neuronal marker expression, whereas glial differentiation was attenuated. A dynamic modulation was also observed, testing the expression of melatonin receptors. In conclusion, our data suggest that melatonin\'s neurogenic differentiation ability can be usefully exploited to obtain neuronal-like differentiated ASCs for potential therapeutic strategies.