Vellosillo Lara, Muñoz Maria Paz, Paíno Carlos Luis
Servicio de Neurobiología-Investigación, IRYCIS, Hospital Universitario Ramón y Cajal, Carretera de Colmenar km 9, 28034, Madrid, Spain.
Histochem Cell Biol. 2017 Nov;148(5):503-515. doi: 10.1007/s00418-017-1588-y. Epub 2017 Jun 15.
Mesenchymal cells cultured from the vasculo-stromal fraction of adipose tissue (ADSC) show adult stem cell characteristics and several groups have claimed generating neural cells from them. However, we have observed that many markers commonly used for the identification of neural cells are spontaneously expressed by ADSC in culture. In the present study, we have examined the expression of characteristic oligodendrocyte molecules in cultured ADSC, aiming to test if myelinating cells could be generated from accessible non-neural adult tissues. In basal growth conditions, rat ADSC spontaneously expressed CNPase, MBP, MOG, protein zero, GAP43, Sox10, and Olig2, as shown by immunocytrochemistry and western blot. A small population of cultured ADSC expressed membrane galactocerebroside (O1 antibody), but no cell stained with O4 antibody. RT-PCR analyses showed the expression of CNPase, MBP, DM20, and low levels of Olig2, Sox10, and Sox2 mRNA by rat ADSC. When rat ADSC were treated with combinations of factors commonly used in neural-inducing media (retinoic acid, dbcAMP, EGF, basic FGF, NT3, and/or PDGF), the number of O1-positive cells changed, but in no case, mRNA expression of Sox10 and Olig2 transcription factors approached CNS oligodendrocyte levels. In co-culture with rat dorsal root ganglion neurons, no sign of axonal myelination by rat ADSC was observed. These studies show that the expression of oligodendrocyte traits by cultured ADSC is not a proof of functional competence as oligodendroglia and suggest that in culture conditions, ADSC acquire intermediate, uncommitted phenotypes.
从脂肪组织的血管基质部分培养的间充质细胞（脂肪来源干细胞）表现出成体干细胞特征，并且有几个研究小组宣称已从它们分化出神经细胞。然而，我们观察到，培养的脂肪来源干细胞可自发表达许多常用于鉴定神经细胞的标志物。在本研究中，我们检测了培养的脂肪来源干细胞中少突胶质细胞特征性分子的表达，旨在验证能否从易于获取的非神经成体组织生成髓鞘形成细胞。在基础生长条件下，通过免疫细胞化学和蛋白质印迹法显示，大鼠脂肪来源干细胞可自发表达2',3'-环核苷酸3'-磷酸二酯酶（CNPase）、髓鞘碱性蛋白（MBP）、髓鞘少突胶质细胞糖蛋白（MOG）、蛋白零、生长相关蛋白43（GAP43）、性别决定区Y框蛋白10（Sox10）和少突胶质细胞转录因子2（Olig2）。一小部分培养的脂肪来源干细胞表达膜半乳糖脑苷脂（O1抗体），但没有细胞被O4抗体染色。逆转录聚合酶链反应（RT-PCR）分析显示，大鼠脂肪来源干细胞表达CNPase、MBP、DM20以及低水平的Olig2、Sox10和Sox2信使核糖核酸（mRNA）。当用神经诱导培养基中常用的因子组合（视黄酸、二丁酰环磷腺苷（dbcAMP）、表皮生长因子（EGF）、碱性成纤维细胞生长因子（bFGF）、神经营养因子3（NT3）和/或血小板衍生生长因子（PDGF））处理大鼠脂肪来源干细胞时，O1阳性细胞数量发生变化，但在任何情况下，Sox10和Olig2转录因子的mRNA表达均未达到中枢神经系统少突胶质细胞水平。与大鼠背根神经节神经元共培养时，未观察到大鼠脂肪来源干细胞有轴突髓鞘形成的迹象。这些研究表明，培养的脂肪来源干细胞表达少突胶质细胞特征并不证明其具有少突胶质细胞的功能能力，并提示在培养条件下，脂肪来源干细胞获得了中间的、未定向的表型。
