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施万细胞可促进神经干细胞分化为神经元的能力,并分泌神经营养因子。

Schwann cells promote the capability of neural stem cells to differentiate into neurons and secret neurotrophic factors.

作者信息

Yu Ziwei, Men Yongzhi, Dong Pin

机构信息

Department of Otolaryngology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China.

出版信息

Exp Ther Med. 2017 May;13(5):2029-2035. doi: 10.3892/etm.2017.4183. Epub 2017 Mar 6.

Abstract

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.

摘要

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。

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