Fang Zhong, Yang Qin, Xiong Wei, Li Guanghui, Xiao Jun, Guo Fengjing, Li Feng, Chen Anmin
Department of Orthopedics, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Feb;30(1):75-80. doi: 10.1007/s11596-010-0113-5. Epub 2010 Feb 14.
Some studies indicate that adipose derived stem cells (ADSCs) can differentiate into adipogenic, chondrogenic, myogenic, and osteogenic cells in vitro. However, whether ADSCs can be induced to differentiate into neural cells in vitro has not been clearly demonstrated. In this study, the ADSCs isolated from the murine adipose tissue were cultured and transfected with the EGFP gene, and then the cells were induced for neural differentiation. The morphology of those ADSCs began to change within two days which developed into characteristics of round cell bodies with several branching extensions, concomitantly expressing EGFP fluorescence. Approximately 60% of the total cell populations were bipolar or multipolar in shape. Some of them appeared to make contact with their neighboring cells. RT-PCR, Western blot and Immunocytochemistry revealed that the expression levels of the markers of neurons and oligodendrocytes such as MAP2, NF-70, Neu N and RIP upon neural induction were increased, but the expression of the special marker of astrocytes, GFAP, was undetectable until 96 h after induction when a small signal was observed. It was concluded that the ADSCs transfected with EGFP possessed the ability to undergo morphologic and phenotypic changes consistent with neural differentiation in vitro. It suggests that these cells might provide an ideal source for further stem cell research with possible therapeutic application for spinal cord injury.
一些研究表明，脂肪来源干细胞（ADSCs）在体外可分化为成脂细胞、成软骨细胞、成肌细胞和成骨细胞。然而，ADSCs在体外是否能被诱导分化为神经细胞尚未得到明确证实。在本研究中，从小鼠脂肪组织分离的ADSCs经培养并用EGFP基因转染，然后诱导细胞进行神经分化。这些ADSCs的形态在两天内开始改变，发展为具有多个分支突起的圆形细胞体特征，同时表达EGFP荧光。约60%的细胞群体呈双极或多极形态。其中一些细胞似乎与相邻细胞有接触。逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法（Western blot）和免疫细胞化学分析显示，神经诱导后神经元和少突胶质细胞标志物如微管相关蛋白2（MAP2）、神经丝蛋白70（NF-70）、神经元核抗原（Neu N）和受体相互作用蛋白（RIP）的表达水平升高，但星形胶质细胞特异性标志物胶质纤维酸性蛋白（GFAP）直到诱导后96小时才检测到表达，此时观察到一个小信号。结论是，转染EGFP的ADSCs具有在体外发生与神经分化一致的形态和表型变化的能力。这表明这些细胞可能为进一步的干细胞研究提供理想来源，并可能对脊髓损伤具有治疗应用价值。
