个体化培养自体黑素细胞移植治疗白癜风

目的 探讨使用个体化培养基进行自体黑素细胞培养移植治疗白癜风的疗效。方法 负压吸疱获取患者正常表皮片，制成细胞悬液，在Hu16黑素细胞选择性培养基中培养。检测黑素细胞分裂时间（DOT）和黑素含量，根据DOT的大小、黑素含量和细胞形态，调整血清、细胞因子浓度及补充内皮素-1，进行个体化黑素细胞培养。经2 ~ 5次传代后收集黑素细胞，白斑区用超脉冲CO2激光磨削后进行黑素细胞移植，随访观察复色效果。结果 共治疗155例稳定期白癜风患者的204处皮损，进行1次移植119例，进行2 ~ 4次移植36例。应用个体化黑素细胞培养后细胞扩增可达50 ~ 80倍。84.80%的皮损复色面积超过50%，其中52.94%的皮损复色面积超过90%，且复色均匀，未见瘢痕及其他不良反应。性别、年龄、病程长短和皮损面积大小对疗效没有影响。节段型白癜风移植疗效好于寻常型白癜风，两组有效率分别为93.62%和82.16%，痊愈率分别为65.96%和49.04%。手臂和腿部的皮损（不包括肘部和膝盖）移植后痊愈率达73.08%，疗效好于躯干、面颈；肢端皮损疗效最差，痊愈率仅为25.93%。结论 个体化培养技术能提高白癜风患者黑素细胞的培养成功率与细胞扩增倍数。体外培养的自体黑素细胞移植治疗稳定期白癜风疗效肯定，用少量供皮区即可治疗大面积皮损，值得临床应用。

Transplantation of individualized cultured autologous melanocytes for the treatment of vitiligo

Objective To evaluate the therapeutic effect of transplantation of autologous melanocytes cultured with individualized medium in vitiligo. Methods Donor skin was obtained by suction blisters from a normally pigmented area of the abdomen of 155 patients with vitiligo. The roof of the blisters was clipped and digested with trypsin, then the suspension of epidermal cells and melanocytes were cultured in Hu16 medium.The cell division time (DOT) and melanin content of cultured melanocytes were measured followed by the adjustment of concentration of fetal calf serum, cytokines and cAMP elevating agents based on the DOT,melanin content and morphology of melanocytes for the individualized culture of melanocytes. After 2 - 5 passages, melanocytes were harvested and inoculated into ultrapluse CO2 laser-denuded lesions. All patients were followed up for at least 6 months. Results One hundred and fifty-five vitiligo patients with 204 lesions were treated with transplantation of autologous melanocytes. Of the 155 patients, 119 received 1 session of transplantation, 36 received 2 to 4 session of transplantation. Cells were expanded by 50 - 80 times in vitro after individualized culture. Repigmentation was more than 50% in 84.8% of these lesions, more than 90% in 52.94% of the lesions. A homogeneous skin color was obtained in repigmented skin, and no scarring or other side effects were observed. No influence was noted on the outcome of transplantation for sex, age, course of disease or lesion size of patients. Segmental vitiligo showed better response than vitiligo vulgaris: the effective rate and cure rate were 93.62% and 65.96% respectively for segmental vitiligo, 82.16% and 49.04% respectively for vitiligo vulgaris. Lesions located on the arms and legs (not including elbows and knees) showed the best response, with a cure rate of 73.08%, whereas acral sites were the most difficult area to repigment, with a cure rate of just 25.93%. Conclusions Individualized culture can significantly increase the success rate of melanocyte culture and expanding times of melanocytes. Transplantation of cultured autologous melanocytes is an effective modality deserving clinical application in the treatment of stable vitiligo, with the advantage of treating large depigmented area with melanocytes from a small donor site.