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脂肪来源干细胞向神经细胞的分化及其在脊髓损伤自体移植中的应用

Neurogenic differentiation from adipose-derived stem cells and application for autologous transplantation in spinal cord injury.

作者信息

Zhao Yong, Jiang Hui, Liu Xin-wei, Chen Jian-Ting, Xiang Liang-Bi, Zhou Da-Peng

机构信息

Department of Orthopedics, Nanfang Hospital of Southern Medical University, Guangzhou, 510515, Guangdong Province, China.

出版信息

Cell Tissue Bank. 2015 Sep;16(3):335-42. doi: 10.1007/s10561-014-9476-3. Epub 2014 Oct 22.

Abstract

Mesenchymal stem cells derived from adipose tissue have the capacity to differentiate into endodermal, mesoderm and ectodermal cell lineages in vitro, which are an ideal engraft in tissue-engineered repair. In this study, mouse adipose-derived stem cells (ADSCs) were isolated from subcutaneous fat. The markers of ADSCs, CD13, CD29, CD44, CD71, CD73, CD90, CD105, CD166, Nestin, GFAP and MAP-2 were detected by immunofluorescence assays. The ADSCs were cultured in cocktail factors (including ATRA, GGF-2, bFGF, PDGF and forskolin) for neurogenic differentiation. The neurogenic cells markers, Nestin, GFAP and MAP-2 were analyzed using immunofluorescence and real-time PCR after dramatic changes in morphology. Neurogenic cells from ADSCs were autologous transplanted into the mouse of spinal cord injury for observation neurogenic cells colonization in spinal cord. The result demonstrated that the mouse ADSCs were positive for the CD13, CD29, CD44, CD71, CD73, CD90, CD105 and CD166 but negative for neurogenic cell markers, MAP-2, GFAP and Nestin. After neurogenic differentiation, the neurogenic cells were positive for neurogenic cell special markers, gene expression level showed a time-lapse increase, and the cells were successful colonized into spinal cord. In conclusion, our research shows that a population of neuronal cells can be specifically generated from ADSCs and that induced cells may allow for participation in tissue-repair.

摘要

源自脂肪组织的间充质干细胞在体外具有分化为内胚层、中胚层和外胚层细胞谱系的能力,是组织工程修复中理想的植入物。在本研究中,从小鼠皮下脂肪中分离出脂肪来源干细胞(ADSCs)。通过免疫荧光分析检测ADSCs的标志物CD13、CD29、CD44、CD71、CD73、CD90、CD105、CD166、巢蛋白、胶质纤维酸性蛋白(GFAP)和微管相关蛋白2(MAP-2)。将ADSCs在鸡尾酒因子(包括全反式维甲酸、生长/分化因子-2、碱性成纤维细胞生长因子、血小板衍生生长因子和福斯高林)中培养以进行神经分化。在形态发生显著变化后,使用免疫荧光和实时定量聚合酶链反应分析神经源性细胞标志物巢蛋白、GFAP和MAP-2。将来自ADSCs的神经源性细胞自体移植到脊髓损伤小鼠中,以观察神经源性细胞在脊髓中的定植情况。结果表明,小鼠ADSCs的CD13、CD29、CD44、CD71、CD73、CD90、CD105和CD166呈阳性,但神经源性细胞标志物MAP-2、GFAP和巢蛋白呈阴性。神经分化后,神经源性细胞的神经源性细胞特异性标志物呈阳性,基因表达水平呈时间依赖性增加,并且这些细胞成功定植于脊髓。总之,我们的研究表明,可以从ADSCs特异性生成一群神经元细胞,并且诱导细胞可能参与组织修复。

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