Xu Chunling, Lu Huayi, Li Fuqiang, Su Guanfang
Department of Ophthalmology, The Second Hospital of Jilin University, Changchun, China.
The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.
J Comput Biol. 2020 Aug;27(8):1329-1336. doi: 10.1089/cmb.2019.0024. Epub 2019 Dec 16.
To explore possible approaches to differentiating rat bone marrow mesenchymal stem cells (BMSCs) into retinal ganglion-like cells and to demonstrate the dynamic changes in protein expression profiles of BMSCs throughout the differentiation. BMSCs were isolated from adult rats and cultured in medium conditioned by neonatal rat retinal cells to induce BMSC differentiation into retinal ganglion-like cells. Immunostaining for neurofilament, nestin, Map2, and Thy1.1 was used to follow the differentiation process. Two types of protein arrays were employed to profile the BMSCs, the differentiated retinal ganglion-like cells, and the primary retinal ganglion cells (RGCs) using the Biomarker Wizard System. After 7 days of culture in conditioned medium, cells showing a neural-cell-like modality appeared. The differentiated retinal ganglion-like cells showed that network-like connections were positive for nestin, neurofilament, Map2, and Thy1.1. In total, 16 marker proteins were highly expressed in both retinal ganglion-like cells and RGCs and no obvious expression was observed in BMSCs. Among them, nine proteins were expressed more highly in RGCs than in retinal ganglion-like cells. BMSCs can be induced to differentiate into retinal ganglion-like cells by neonatal rat retinal cells, and the induced cells show protein profiles resembling those of isolated RGCs.
探索将大鼠骨髓间充质干细胞(BMSCs)分化为视网膜神经节样细胞的可能方法,并展示BMSCs在整个分化过程中蛋白质表达谱的动态变化。从成年大鼠中分离出BMSCs,并在新生大鼠视网膜细胞条件培养基中培养,以诱导BMSCs分化为视网膜神经节样细胞。使用神经丝、巢蛋白、Map2和Thy1.1的免疫染色来跟踪分化过程。采用两种蛋白质芯片,使用生物标志物向导系统对BMSCs、分化的视网膜神经节样细胞和原代视网膜神经节细胞(RGCs)进行蛋白质谱分析。在条件培养基中培养7天后,出现了呈现神经细胞样形态的细胞。分化的视网膜神经节样细胞显示,网络状连接对巢蛋白、神经丝、Map2和Thy1.1呈阳性。共有16种标志物蛋白在视网膜神经节样细胞和RGCs中高表达,而在BMSCs中未观察到明显表达。其中,9种蛋白在RGCs中的表达高于视网膜神经节样细胞。新生大鼠视网膜细胞可诱导BMSCs分化为视网膜神经节样细胞,诱导后的细胞显示出与分离的RGCs相似的蛋白质谱。