毛乳头细胞的培养与鉴定

Culture and identification of hair dermal papilla cells

目的 :建立毛乳头细胞的培养方法 ,以进一步研究毛乳头细胞在毛发再生中的作用和意义 .方法 :分离得到完整毛囊 ,依次采用Dispase酶和胶原酶消化得到毛乳头 ,再进行毛乳头细胞的培养 .结果 :成功进行了毛乳头细胞的原代和传代培养 ,所获细胞经细胞化学染色证实细胞具有明显的异染特性 .毛乳头贴壁后 5～ 6d周围即可见细胞长出 ,细胞生长快 ,1 2～ 1 6d左右细胞即可融合 .在倒置显微镜下细胞呈成纤维细胞样 ,有聚集生长特性 ,至少可以传 1 5代 .冻存后复苏细胞贴壁率在 80 %以上 .结论 :该方法是体外培养毛乳头细胞的理想方法 ,操作较为简单 ,降低了工作强度 ,所获细胞纯度高 ,可以长期传代。

AIM: To establish a culture method of the hair dermal papilla cells (DPc) and to further investigate the role of DPcs in hair regeneration. METHODS: The whole hair follicle was separated, and digested by dispase and collagenase, and the hair papilla was cultured in the medium. RESULTS: DPcs with hair papilla separated by dispase and collagenase were cultured successfully. The cells could be identificated by cytochemistry staining. Fibroblast like under invert microscope, the cells possessed aggregation properties. After the hair papilla adhibited for 5-6 d, the DPcs grew around it very quickly, and overspread after 12-16 d. It could be subcultured to 15 passages. The recovery attachment efficiency of DPcs in less than passage 10 was more than 80%. CONCLUSION: The method is simple and effective in DPcs culture. The acquired cells are very pure and can be subcultured or frozen for a long time.