Molecular Biology Laboratory, Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Kunming 650118, Yunnan Province, China.
Neural Regen Res. 2012 Dec 5;7(34):2645-52. doi: 10.3969/j.issn.1673-5374.2012.34.001.
LIM homeobox transcription factor 1a (Lmx1a) has the capacity to initiate the development program of neuronal cells and promote the differentiation of embryonic stem cells into dopaminergic neurons. In this study, rhesus adipose stem cells were infected with recombinant adenovirus carrying the Lmx1a gene and co-cultured with embryonic rat neural stem cells. Cell differentiation was induced using sonic hedgehog and fibroblast growth factor-8. Immunofluorescence staining showed that cells were positive for neuron-specific enolase and β-tubulin III. Reverse transcription-PCR results demonstrated that rhesus adipose stem cells were not only positive for neuron-specific enolase and β-tubulin III, but also positive for the dopaminergic neuron marker, tyrosine hydroxylase, neurofilament, glial cell line-derived neurotrophic factor family receptor α2 and nuclear receptor related factor 1. The number of Lmx1a gene-infected cells expressing the dopaminergic neuron marker was substantially greater than the number of cells not infected with Lmx1α gene. These results suggest that Lmx1a-mediated regulation combined with the strategy of co-culture with neural stem cells can robustly promote the differentiation of rhesus adipose stem cells into dopaminergic neurons.
LIM 同源盒转录因子 1a（Lmx1a）具有启动神经元细胞发育程序和促进胚胎干细胞向多巴胺能神经元分化的能力。在这项研究中，恒河猴脂肪干细胞被携带 Lmx1a 基因的重组腺病毒感染，并与胚胎大鼠神经干细胞共培养。使用 sonic hedgehog 和纤维母细胞生长因子-8 诱导细胞分化。免疫荧光染色显示细胞对神经元特异性烯醇化酶和 β-微管蛋白 III 呈阳性。逆转录-PCR 结果表明，恒河猴脂肪干细胞不仅对神经元特异性烯醇化酶和 β-微管蛋白 III 呈阳性，而且对多巴胺能神经元标志物酪氨酸羟化酶、神经丝、胶质细胞源性神经营养因子家族受体 α2 和核受体相关因子 1 也呈阳性。表达多巴胺能神经元标志物的 Lmx1a 基因感染细胞的数量明显多于未感染 Lmx1α 基因的细胞数量。这些结果表明，Lmx1a 介导的调节与与神经干细胞共培养的策略相结合，可以强有力地促进恒河猴脂肪干细胞向多巴胺能神经元分化。
