EdU体外标记毛囊外根鞘干细胞的实验研究  

Experimental Study of EdU in Vitro Labeling of Hair Follicle Outer Root Sheath Stem Cells

作　　者：王海涛[1] 朱梓赫 王丽[1] 王泽鹏[1] 李树伟[1] 

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：《现代生物医学进展》2017年第8期1425-1429,1451,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31560685);塔里木大学硕士创新项目(TDGRI201521)

摘　　要：目的:探寻一种简单有效的追踪定位毛囊干细胞的分裂增殖标记的方法,为后期毛囊干细胞增殖分化迁移机制的研究打下基础。方法:体外无菌条件下获取昆明乳鼠触须毛囊外根鞘,以5%FCS+MEM作为基础培养液,添加4种不同浓度EdU(5-Ethynyl-2’-deoxyuridine)细胞标记物,进行毛囊外根鞘再生培养,冰冻切片,H.E染色,使用Key Fluor488对EdU所标记的再生细胞进行检测。结果:H.E染色和EdU标记染色结果发现EdU细胞标记物浓度越高,毛囊外根鞘再生形态越差,同时当EdU浓度为20μmol/L时,毛囊外根鞘增殖细胞标记效果是最好的。结论:EdU细胞标记物对毛囊外根鞘中的干细胞增殖活性具有一定的毒副作用,可能会影响细胞的正常生理代谢功能,浓度为20μmol/L EdU可以有效的标记毛囊干细胞增殖,并对毛囊再生结构影响较小。Objective： To explore a simple and effective method to track the proliferation and differentiation of hair follicle stem cells, and to lay a foundation for the study of the mechanism of proliferation and differentiation of hair follicle stem cells in the later stage. Methods： Vibrissa follicle outer root sheath of KM mouse was collected under sterile conditions 5 % FCS＋MEM was used as basic culture liquid with 4 different concentrations of EdU （5-Ethynyl-2＇-deoxyuridine） as cells in the marker to the outer root sheath of the hair follicle regeneration culture and frozen section, after H.E. staining, the regenerative cells labeled with EdU were detected using KeyFluor488. Results： According to the results of H.E staining and the EdU labeling and staining, hair follicle outer root sheath regeneration morphological is worse when the cell marker concentration is high, and the marker effect of hair follicle outer root sheath cell proliferation is the best when the EdU concentration of 20 μmol/L. Conclusion： EdU cell markers of hair follicle outer root sheath stem cell proliferation activity has certain side effects affect on cell＇s normal physiological function, the concentration of 20 μmol/L EdU can be effectively labeled by the hair follicle stem cell proliferation, and on hair follicle regeneration structure effect is small.

关 键 词：触须毛囊 毛囊干细胞 Ed U 细胞增殖 

分 类 号：Q-31[生物学] Q593