Eriksson Cecilia, Wictorin Klas
Wallenberg Neuroscience Center, Division of Neurobiology, Lund University, Lund, Sweden.
Exp Neurol. 2003 Dec;184(2):1058-63. doi: 10.1016/j.expneurol.2003.08.014.
Attached glial-like cell cultures were established from the lateral and medial ganglionic eminences (LGE and MGE) and from the neocortex (Cx) of E13.5 mouse embryos, and expanded over four to five passages under epidermal growth factor (EGF) stimulation. Following removal of EGF and serum, we analysed the generation of neurons and glial cells within the cultures. Significant numbers of betaIII-tubulin-positive neurons were generated in both the LGE (about 7% of total cell numbers) and the MGE (around 2%). However, only few betaIII-tubulin-positive cells with neuronal morphologies were detected in the differentiated Cx cultures. The newly formed neurons were to a large extent GABAergic, and many of the MGE-derived, but not the LGE-derived, cells expressed the MGE-marker NKX2.1. Most cells in all cultures still appeared astroglial-like, expressing glial fibrillary acidic protein (GFAP), but in addition, CNPase-positive cells with oligodendroglial morphologies were present in the MGE (0.68%), and, to a lesser extent (0.2%), in the LGE cultures. The present results demonstrate that cells of expanded glial cultures from both the LGE and MGE can give rise to significant and, to a certain extent, region-specific neuronal and glial cell types under differentiating conditions.
从E13.5小鼠胚胎的外侧和内侧神经节隆起(LGE和MGE)以及新皮质(Cx)建立附着的类神经胶质细胞培养物,并在表皮生长因子(EGF)刺激下传代培养四到五代。去除EGF和血清后,我们分析了培养物中神经元和神经胶质细胞的生成情况。在LGE(约占细胞总数的7%)和MGE(约2%)中均产生了大量βIII-微管蛋白阳性神经元。然而,在分化的Cx培养物中仅检测到少数具有神经元形态的βIII-微管蛋白阳性细胞。新形成的神经元在很大程度上是γ-氨基丁酸能的,并且许多源自MGE而非LGE的细胞表达MGE标记物NKX2.1。所有培养物中的大多数细胞仍表现出星形胶质样,表达胶质纤维酸性蛋白(GFAP),但此外,MGE中存在具有少突胶质细胞形态的CNPase阳性细胞(0.68%),在LGE培养物中的比例较低(0.2%)。目前的结果表明,来自LGE和MGE的扩增神经胶质培养细胞在分化条件下可以产生大量且在一定程度上具有区域特异性的神经元和神经胶质细胞类型。