Department of Psychiatry and the Nina Ireland Laboratory of Developmental Neurobiology, University of California San Francisco, San Francisco, CA, USA.
PLoS One. 2013 May 1;8(5):e61956. doi: 10.1371/journal.pone.0061956. Print 2013.
The medial ganglionic eminence (MGE) is an embryonic forebrain structure that generates the majority of cortical interneurons. MGE transplantation into specific regions of the postnatal central nervous system modifies circuit function and improves deficits in mouse models of epilepsy, Parkinson's disease, pain, and phencyclidine-induced cognitive deficits. Herein, we describe approaches to generate MGE-like progenitor cells from mouse embryonic stem (ES) cells. Using a modified embryoid body method, we provided gene expression evidence that mouse ES-derived Lhx6(+) cells closely resemble immature interneurons generated from authentic MGE-derived Lhx6(+) cells. We hypothesized that enhancers that are active in the mouse MGE would be useful tools in detecting when ES cells differentiate into MGE cells. Here we demonstrate the utility of enhancer elements [422 (DlxI12b), Lhx6, 692, 1056, and 1538] as tools to mark MGE-like cells in ES cell differentiation experiments. We found that enhancers DlxI12b, 692, and 1538 are active in Lhx6-GFP(+) cells, while enhancer 1056 is active in Olig2(+) cells. These data demonstrate unique techniques to follow and purify MGE-like derivatives from ES cells, including GABAergic cortical interneurons and oligodendrocytes, for use in stem cell-based therapeutic assays and treatments.
内侧神经节隆起（MGE）是一种胚胎前脑结构，它产生了大部分皮质中间神经元。将 MGE 移植到出生后中枢神经系统的特定区域可以改变回路功能，并改善癫痫、帕金森病、疼痛和苯环己哌啶引起的认知缺陷的小鼠模型的缺陷。在此，我们描述了从小鼠胚胎干细胞（ES 细胞）中生成 MGE 样祖细胞的方法。使用改良的胚状体方法，我们提供了基因表达证据，证明从小鼠 ES 细胞衍生的 Lhx6(+)细胞与源自真实 MGE 衍生的 Lhx6(+)细胞的未成熟中间神经元非常相似。我们假设在小鼠 MGE 中活跃的增强子将是检测 ES 细胞何时分化为 MGE 细胞的有用工具。在这里，我们展示了增强子元件 [422（DlxI12b）、Lhx6、692、1056 和 1538] 作为在 ES 细胞分化实验中标记 MGE 样细胞的工具的有用性。我们发现，增强子 DlxI12b、692 和 1538 在 Lhx6-GFP(+)细胞中活跃，而增强子 1056 在 Olig2(+)细胞中活跃。这些数据证明了独特的技术，可以从 ES 细胞中追踪和纯化 MGE 样衍生物，包括 GABA 能皮质中间神经元和少突胶质细胞，用于基于干细胞的治疗性测定和治疗。
