小鼠触须毛囊外根鞘再生的体外培养液优化及α-SMA的表达鉴定  

Optimization Medium of Culturing the Mice Whisker Outer Root Sheath in Vitro and Identification of Expression of α-SMA

作　　者：唐瑶 邓梅成 王海涛[1] 付灵慧 李树伟 Tang Yao Deng Meicheng Wang Haitao Fu Linghui Li Shuwei(Key Laboratory of Protection and Utilization of Biological Resource in Tarim Basin of Xinjiang Production ＆ Construction Corps/College of Life Science, Tarim University, Alar, Xinjiang 84330)

机构地区：[1]塔里木盆地生物资源保护利用兵团重点实验室/塔里木大学生命科学学院,新疆阿拉尔843300

出　　处：《塔里木大学学报》2016年第2期1-7,共7页Journal of Tarim University

基　　金：国家自然科学基金(31560685);国家级大学生创新项目(201410757009);塔里木大学校级大学生创新项目(2014010);塔里木大学研究生科研创新校级指导项目(TDGRI201521)

摘　　要：【目的】为建立小鼠毛囊体外培养模型,筛选适合小鼠触须毛囊外根鞘离体培养的培养液,并进行α-SMA(α-平滑肌肌动蛋白)表达的免疫荧光鉴定。【方法】选择出生5 d内的乳鼠断颈法处死,利用显微解剖技术,分离触须毛囊外根鞘。并分别培养于无血清、含5%FCS、10%FCS的DMEM、MEM共计6种不同培养液中,在37℃,5%CO2培养箱中培养4 d后,制作冰冻切片并进行H.E.染色,在显微镜下观察乳鼠触须毛囊外根鞘的再生情况,筛选培养液。在最适培养液中培养乳鼠触须毛囊外根鞘,利用α-SMA抗体进行乳鼠触须毛囊外根鞘再生部位α-SMA表达的荧光鉴定。【结果】观察到小鼠触须毛囊外根鞘中段再生的初期结构特征,并且5%FCS MEM培养液是最适合小鼠触须毛囊外根鞘的体外培养液,免疫荧光检测发现α-SMA在毛囊外根鞘再生部位有表达。【结论】5%FCS MEM培养液是比较适合小鼠触须毛囊外根鞘的体外培养液,α-SMA的表达可以作为毛囊再生的检测指标之一。[ Objective ] To establish the mouse follicle in vitro culture model, optimized the culture medium of the mice outer root sheath, and detected α- SMA （ α- smooth muscle actin） expression by immunofluorescence. [ Methods ] Newborn mice were sacri- riced from cervical, isolated mice outer root sheath with microsurgical dissection, and cultured in 6 kinds of medium such as serum - free, 5% FCS （fetal calf serum） and 10% FCS DMEM or MEM respectively. Put them in 5% CO2 incubator at 37℃ for4 days, em- bedding and frozen section, and then H.E. staining. The mice outer root sheath regeneration were observed under the microscope ob- servation to select the best culture medium. Rat vibrissa follicle outer root sheath were cultured in the optimum culture liquid, and Im-munoflourescence analysis the newborn mice outer root sheath regeneration with ot -SMA antibody. [ Results ] The early stage of the mice follicle outer root sheath regeneration were observed. 5% FCS MEM medium were suitable culture medium for the mice outer root sheath regeneration. Immunofluorescence analysis found that α-SMA expressed at the region of regeneration. [ Conclusion] 5% MEM medium were more suitable for culturing mice outer root sheath in vitro, and α - SMA expression could be used as indicator to detect the whisker follicle regeneration.

关 键 词：小鼠毛囊 触须 外根鞘 免疫荧光 α-SMA抗体 

分 类 号：Q954.539[生物学—动物学]