Chen Song, Zhang Wei, Wang Ji-Ming, Duan Hong-Tao, Kong Jia-Hui, Wang Yue-Xin, Dong Meng, Bi Xue, Song Jian
Tianjin Eye Hospital, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin Eye Institute, Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300020, China.
Int J Ophthalmol. 2016 Jan 18;9(1):41-7. doi: 10.18240/ijo.2016.01.07. eCollection 2016.
To investigate whether umbilical cord human mesenchymal stem cell (UC-MSC) was able to differentiate into neural stem cell and neuron in vitro.
The umbilical cords were obtained from pregnant women with their written consent and the approval of the Clinic Ethnics Committee. UC-MSC were isolated by adherent culture in the medium contains 20% fetal bovine serum (FBS), then they were maintained in the medium contain 10% FBS and induced to neural cells in neural differentiation medium. We investigated whether UC-MSC was able to differentiate into neural stem cell and neuron in vitro by using flow cytometry, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunofluorescence (IF) analyzes.
A substantial number of UC-MSC was harvested using the tissue explants adherent method at about 2wk. Flow cytometric study revealed that these cells expressed common markers of MSCs, such as CD105 (SH2), CD73 (SH3) and CD90. After induction of differentiation of neural stem cells, the cells began to form clusters; RT-PCR and IF showed that the neuron specific enolase (NSE) and neurogenic differentiation 1-positive cells reached 87.3%±14.7% and 72.6%±11.8%, respectively. Cells showed neuronal cell differentiation after induced, including neuron-like protrusions, plump cell body, obviously and stronger refraction. RT-PCR and IF analysis showed that microtubule-associated protein 2 (MAP2) and nuclear factor-M-positive cells reached 43.1%±10.3% and 69.4%±19.5%, respectively.
Human umbilical cord derived MSCs can be cultured and proliferated in vitro and differentiate into neural stem cells, which may be a valuable source for cell therapy of neurodegenerative eye diseases.
研究脐带间充质干细胞（UC-MSC）在体外是否能够分化为神经干细胞和神经元。
在获得孕妇书面同意并经临床伦理委员会批准后获取脐带。通过在含有20%胎牛血清（FBS）的培养基中贴壁培养分离UC-MSC，然后将其置于含有10%FBS的培养基中，并在神经分化培养基中诱导分化为神经细胞。我们通过流式细胞术、逆转录聚合酶链反应（RT-PCR）和免疫荧光（IF）分析来研究UC-MSC在体外是否能够分化为神经干细胞和神经元。
使用组织块贴壁法在约2周时收获了大量的UC-MSC。流式细胞术研究显示这些细胞表达间充质干细胞的常见标志物，如CD105（SH2）、CD73（SH3）和CD90。在诱导神经干细胞分化后，细胞开始形成细胞团；RT-PCR和IF显示神经元特异性烯醇化酶（NSE）和神经源性分化1阳性细胞分别达到87.3%±14.7%和72.6%±11.8%。诱导后细胞呈现神经元细胞分化，包括神经元样突起、饱满的细胞体、明显且较强的折光性。RT-PCR和IF分析显示微管相关蛋白2（MAP2）和核因子-M阳性细胞分别达到43.1%±10.3%和69.4%±19.5%。
人脐带源间充质干细胞可在体外培养和增殖并分化为神经干细胞，这可能是神经退行性眼病细胞治疗的宝贵来源。
