组织块法与酶消化法培养大鼠毛囊干细胞的比较

摘要

BACKGROUND:Hair folicle stem cels have been confirmed to have stronger proliferative ability than interfolicular epidermal stem cels, which have been an issue of concern in seed cel research. OBJECTIVE:To compare the biological characteristics of rat hair folicle stem cels cultured by tissue explant method and enzymatic digestion method. METHODS: Under stereomicroscope, hair folicles were isolated from the rat whiskers, and then tissue explant method and two-step enzymatic digestion method were employed to culture hair folicle stem cels. Cels were purified using repeated differential adhesion method, and cel growth and morphology were observed periodicaly. Flow cytometry was used to detect the expression of CD34 and β1 integrin in passage 3 hair folicle stem cels. RESULTS AND CONCLUSION:Cels cultured by two-step enzymatic digestion method grew faster with more amount than those cultured by tissue explants method. Flow cytometry showed that the expressions of PE-CD34 and FITC-β1 were (39.52±19.57)% and (93.46±4.73)% for the two-step enzymatic digestion group, and (19.20±11.53)% and (363.57±14.42)% for the tissue explant method, respectively. There was a significant difference between the two methods. In conclusion, these two methods are able to culture high-activity hair folicle stem cels, which can be chosen according to different experimental requirements.%背景：研究证实毛囊干细胞比毛囊间表皮干细胞更有增生能力，近年来受到广泛关注，成为种子细胞的研究热点。目的：比较组织块法和两步酶法培养大鼠毛囊干细胞的生物学特性。方法：体式显微镜下分离大鼠触须部的毛囊，分别用组织块法和两步酶法培养毛囊干细胞，利用反复差速贴壁法纯化细胞，定期观察细胞生长状况及形态，流式细胞仪检测第3代毛囊干细胞CD34、β1整合素的表达。结果与结论：两步酶法获得的细胞生长速度快，获得的细胞量多，而组织块法获得的细胞生长速度较慢，获得的细胞量也少。流式细胞仪分析显示酶消化法培养组 PE-CD34、FITC-β1整合素的表达分别为(39.52±19.57)%和(93.46±4.73)%，组织块法培养组相应为(19.20±11.53)%和(363.57±14.42)%，两组间差异有显著性意义(P <0.05)。总的来说，两种方法均能培养出实验所需毛囊干细胞，可根据不同实验需求选择恰当的培养方法。