[1]王永飞,赵建华,晁瑞,等.DCs和NSPCs共培养促进NSPCs增殖及其机制的初步研究[J].第三军医大学学报,2012,34(05):373-377.
 Wang Yongfei,Zhao Jianhua,Chao Rui,et al.Dendritic cells promote neural stem/progenitor cells proliferation in vitro when cocultured[J].J Third Mil Med Univ,2012,34(05):373-377.
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DCs和NSPCs共培养促进NSPCs增殖及其机制的初步研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第05期
页码:
373-377
栏目:
论著
出版日期:
2012-03-15

文章信息/Info

Title:
Dendritic cells promote neural stem/progenitor cells proliferation in vitro when cocultured
作者:
王永飞赵建华晁瑞陈太邦郭乔楠
第三军医大学大坪医院野战外科研究所脊柱外科;第三军医大学西南医院病理学研究所
Author(s):
Wang Yongfei Zhao Jianhua Chao Rui Chen Taibang Guo Qiaonan
Department of Spinal Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042; Institute of Pathology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
神经干细胞树突状细胞神经营养素-3酪氨酸激酶受体C
Keywords:
neural stem/progenitor cells dendritic cells neurotrophin-3 tyrosine kinase receptors C
分类号:
R329-33;R329.28;R338.1
文献标志码:
A
摘要:
目的      观察体外Transwell共培养系统中树突状细胞(dendritic cells,DCs)对神经干/前体细胞(neural stem/progenitor cells,NSPCs)增殖的影响,初步探讨神经营养因子-3(neurotrophin-3, NT-3)在DCs调控NSPCs中的作用机制。      方法       根据是否采用Transwell小室将DCs和NSPCs共培养分为分隔培养(DCs/NSPCs组)和混合培养(DCs+NSPCs组),ELISA法测定DCs组、NSPCs组、DCs+NSPCs组、DCs/NSPCs组共培养48 h上清中的NT-3的含量。为观察NT-3在DCs调控NSPCs中的作用及可能的机制,成球法检测NSPCs组、NSPCs/DCs+K252a组、NSPCs+DCs组、NSPCs+NT-3组的NSPCs增殖,免疫细胞荧光法和Western blot法检测各组NSPCs表面酪氨酸激酶受体C(TrkC)的表达。      结果       ELISA实验结果显示,Transwell共培养48 h上清NT-3的含量,NSPCs/DCs组较DCs组和NSPCs组明显升高(P<0.05)。镜下观察、测量结果显示NSPCs/DCs组和NSPCs+NT-3组神经球数量增多,平均直径明显增大(P<0.05)。免疫细胞荧光和Western blot检测结果表明,NSPCs+DCs组和NSPCs+NT-3组中NSPCs的TrkC表达较NSPCs组和NSPCs/DCs+K252a组高(P<0.05)。      结论       体外DCs与NSPCs共培养,促进NSPCs增殖、NT-3的分泌及TrkC的表达,NT-3可能是通过TrkC受体参与NSPCs增殖的调控。
Abstract:
Objective      To investigate the effect of dendritic cells (DCs) coculture on the proliferation of neural stem/progenitor cells (NSPCs) in vitro, and explore the role of TrkC receptor and neurotrophin-3 (NT-3) in the process.       Methods      According to whether to adopt the Transwell chamber, the DCs and the NSPCs were co-cultured and divided into separate culture (DCs/NSPCs group) and mixed culture (DCs+NSPCs group, without the use of Transwell chamber). ELISA was used to detect neurotrophin-3 (NT-3) protein in the supernatant in 48 h after culture in NSPCs+DCs group (mixed culture), DCs group, NSPCs group, and NSPCs/DCs group (only cells separated by Transwell chamber). Tyrosine kinase receptors C (TrkC) proteins on the surface of NSPCs were detected by immunofluorescence staining and Western blotting analysis in NSPCs group, NSPCs/DCs+K252a group (TrKC blocker), NSPCs+DCs group, and NSPCs+NT-3 group.       Results      The levels of NT-3 in the supernatant were significantly higher in 48 h after culture in NSPCs/DCs group than in DCs group and NSPCs group (P<0.05). And microscopy showed significantly increases in the number and the average diameter of neurospheres in NSPCs/DCs group (P<0.05). Immunofluorescence staining and Western blotting indicated that the expression level of TrkC were significantly higher in NSPCs/DCs group and NSPCs+DCs group than in NSPCs group and NSPCs/DCs+K252a group (P<0.05).       Conclusion      DCs in coculture promotes NSPCs proliferation in vitro, which might be through TrkC-NT-3 signal pathway.

参考文献/References:

王永飞, 赵建华, 晁瑞, 等. DCs和NSPCs共培养促进NSPCs增殖及其机制的初步研究[J].第三军医大学学报,2012,34(5):373-377.

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更新日期/Last Update: 2012-03-01