人毛囊外根鞘及毛乳头细胞的迁移和生长观察

Migration and Growth of Cells from Human Hair Follicle Outer Root Sheath and Dermal Papilla Cells

目的连续观察成人头皮毛囊外根鞘(ORS)和毛乳头(DP)细胞的生长及形态学变化。方法利用两步酶消化获得正常人头皮毛囊的ORS和DP,采用添加HKGS和HMGS的K-SFM进行体外培养,倒置显微镜下连续观察毛囊ORS和DP的迁移和生长。结果将单个毛囊置于预先涂布血清的培养板2h,然后添加少量培养基,毛囊黏附于培养板的几率增加,数小时后,即可见细胞从ORS及DP处迁移生长,以角质形成细胞(KCs)、成纤维细胞(FBs)为主,黑素细胞(MCs)较少。并且,ORS和DP的迁出和生长有明显不同。如果培养基过多,则多数毛囊悬浮,无细胞迁出,最终死亡。结论预先涂布血清和开始仅添加少量培养基,有利于毛囊黏附,随后细胞迁移生长。

Objective Continuous observation of adult scalp hair follicle outer root sheath （ORS） and dermal papilla （DP） cells growth and morphological changes. Methods Single hair follicles were completly isolated by collagenase IV and dispase Ⅱ Growth and migration of adult scalp hair follicle ORS and DP cells were continuously observed under inverted microscope. Results Single hair follicle were put onto the dish that serum-coated in advance cultured for 2 hours, then added a small amount of culture medium, the hair follicle sticking probability would increase and some cells migrated from hair follicle ORS and DP ceils some hours later, mainly being keratinocytes （ KCs）, fibroblasts （FBs） and fewer melanocytes （MCs）. Besides, there were obviously differences in cells growth and migration between hair follicle ORS and DP ceils. If added culture medium excessively, most hair follicle would suspend readily, no cells migrated and died finally. Conclusion Serum-coated in advance and a small amount of culture medium at the very start can promote hair follicle to adhere, then cells started to migrate and grow.