Yu Ziwei, Men Yongzhi, Dong Pin
Department of Otolaryngology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China.
Exp Ther Med. 2017 May;13(5):2029-2035. doi: 10.3892/etm.2017.4183. Epub 2017 Mar 6.
The present study investigated whether co-culturing Schwann cells (SCs) with neural stem cells (NSCs) improves viability, direction of differentiation and secretion of brain-derived neurotrophic factor (BDNF) and glial cell-derived neurotrophic factor (GDNF) in NSCs. The three groups assessed were as follows: SCs, NSCs, and a co-culture of SCs and NSCs. Cellular morphological changes were observed under an inverted phase contrast microscope and quantified. Cells were identified by immunofluorescence staining: S100 for SCs, Nestin for NSCs, microtubule associated protein (Map) 2 and NeuN for neurons and glial fibrillary acidic protein for astrocytes. Cell viability was evaluated by MTT assay. Secretion of BDNF and GDNF was quantified; mRNA expression was quantified by reverse transcription-quantitative polymerase chain reaction. The majority of NSCs in the co-cultured group differentiated into neurons. The cell survival rate of the co-culture group was significantly higher than the other groups on days 3, 5 and 10 (P<0.01). The secretion of BDNF in the co-culture group was significantly higher than NSCs on days 3, 5 and 7 (P<0.05), while the amount of GDNF in co-culture was significantly higher than both NSCs and SCs on day 1 (P<0.05). BDNF and GDNF gene expression in the co-culture group was significantly higher than SCs (P<0.01). Gene expression of Map2 in co-culture group was also significantly higher than both NSC and SC groups (P<0.01). Therefore, co-cultured SCs and NSCs promote differentiation of NSCs into neurons and secrete higher levels of neurotropic factors including BDNF and GDNF.
本研究调查了雪旺细胞（SCs）与神经干细胞（NSCs）共培养是否能提高NSCs的活力、分化方向以及脑源性神经营养因子（BDNF）和胶质细胞源性神经营养因子（GDNF）的分泌。评估的三组如下：SCs组、NSCs组以及SCs与NSCs的共培养组。在倒置相差显微镜下观察细胞形态变化并进行量化。通过免疫荧光染色鉴定细胞：SCs用S100染色，NSCs用巢蛋白染色，神经元用微管相关蛋白（Map）2和神经元核抗原（NeuN）染色，星形胶质细胞用胶质纤维酸性蛋白染色。通过MTT法评估细胞活力。对BDNF和GDNF的分泌进行量化；通过逆转录定量聚合酶链反应对mRNA表达进行量化。共培养组中的大多数NSCs分化为神经元。在第3、5和10天，共培养组的细胞存活率显著高于其他组（P<0.01）。在第3、5和7天，共培养组中BDNF的分泌显著高于NSCs组（P<0.05），而在第1天，共培养组中GDNF的含量显著高于NSCs组和SCs组（P<0.05）。共培养组中BDNF和GDNF基因表达显著高于SCs组（P<0.01）。共培养组中Map2的基因表达也显著高于NSC组和SCs组（P<0.01）。因此，共培养的SCs和NSCs促进NSCs向神经元分化，并分泌更高水平的包括BDNF和GDNF在内的神经营养因子。
