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小鼠CD4~+T细胞外泌体分离及鉴定
引用本文:张伟鑫,曾仲.小鼠CD4~+T细胞外泌体分离及鉴定[J].中国比较医学杂志,2017,27(10):60-64.
作者姓名:张伟鑫  曾仲
作者单位:昆明医科大学第一附属医院器官移植中心, 昆明 650000,昆明医科大学第一附属医院器官移植中心, 昆明 650000
基金项目:云南省卫生科技计划基金资助项目(2014NS155)。
摘    要:目的探讨CD4~+T细胞是否具有分泌外泌体的能力,以及CD4~+T细胞外泌体参与炎症及肿瘤免疫过程的可能。方法应用断颈处死的方法取出小鼠的脾脏,用磁珠分选的方法从小鼠脾脏细胞中分选出CD4~+T细胞,并尝试从培养基上清液中提取CD4~+T细胞的外泌体,在透射电镜下观察,后用PCR技术分析miR-23a、miR-214的表达情况。结果在透射电镜下能够观察到提取出的CD4~+T细胞外泌体,且用PCR技术能检测到小鼠CD4~+T细胞外泌体中miR-23a、miR-214这两个炎症及肿瘤相关的miRNA的表达。结论小鼠CD4~+T细胞有分泌外泌体的能力,且为CD4~+T细胞外泌体参与炎症及肿瘤相关过程提供了可能。

关 键 词:CD4+T细胞  外泌体  分离  鉴定  miRNA
收稿时间:2017/2/28 0:00:00

Isolation and identification of exosomes of CD4+T cells in mice
ZHANG Wei-xin and ZENG Zhong.Isolation and identification of exosomes of CD4+T cells in mice[J].Chinese Journal of Comparative Medicine,2017,27(10):60-64.
Authors:ZHANG Wei-xin and ZENG Zhong
Institution:Organ Transplantation Center, The First Affiliated Hospital of Kunming Medical University, Kunming 650000, China and Organ Transplantation Center, The First Affiliated Hospital of Kunming Medical University, Kunming 650000, China
Abstract:Objective To explore whether CD4+T cells in mice can secrete exosomes and the possibility of exosomes to participate in the inflammation and immune response process. Methods Spleen samples were taken from mice, and CD4+T cells were isolated from the spleen tissue using magnetic bead separation method. Exosomes of CD4+ T cells were extracted from the supernatant and observed under transmission electron microscope. PCR assay was used to detect the expression of miR-23a and miR-214. Results Exosomes were observed under elecron microscope,and the expression of miR-23a and miR-214 were detected by PCR assay.Conclusions CD4+T cells in mice can secrete exosomes,which offer possibility to participate in the inflammation and immune response.
Keywords:CD4+T cells  Exosome  Isolation  Identication  MiRNA
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