中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (9): 618-622.

• 论著 • 上一篇    下一篇

黑素细胞个体化培养体系的建立及应用

洪为松1,傅丽芳2,尉晓冬1,许爱娥3   

  1. 1. 杭州市第三人民医院
    2.
    3. 安徽医科大学附属杭州市第三人民医院皮肤科
  • 收稿日期:2010-12-06 修回日期:2011-02-24 出版日期:2011-09-15 发布日期:2011-08-31
  • 通讯作者: 许爱娥 E-mail:xuaiehz@msn.com
  • 基金资助:

    浙江省科技厅重大科技专项国际合作项目;杭州市科技局

Establishment and application of individualized culture system for melanocytes

  • Received:2010-12-06 Revised:2011-02-24 Online:2011-09-15 Published:2011-08-31

摘要:

目的 建立有效的黑素细胞个体化培养实验方案,并观察其治疗大面积白癜风的疗效。方法 Hu16培养基体外培养黑素细胞,应用黑素细胞的分裂时间(DOT)、黑素含量(M)、黑素制造量(MP)和树突数作为黑素细胞生物学活性的指标。通过调整Hu16培养基成分进行个体化培养,以获得生物学活性较好的黑素细胞。皮损区应用超脉冲二氧化碳激光磨削后进行黑素细胞移植,随访观察疗效。结果 共培养了10例稳定期节段型白癜风患者黑素细胞,其中6例黑素细胞(第1 ~ 7代)的DOT较小,M、MP相对稳定,判定黑素细胞生长处于开始或生长的阶段,适合进行细胞移植。另4例黑素细胞从第3代开始DOT较大,M、MP和细胞树突数不稳定,通过调整培养基成分,使黑素细胞生长处于生长阶段,并用于移植。经12个月随访后,7例复色面积超过90%,3例复色面积为70% ~ 80%,平均移植面积达(116.8 ± 75.6) cm2。结论 通过调整黑素细胞培养基成分进行个体化培养可获得生物学活性较好的黑素细胞,用于治疗稳定期节段型白癜风的大面积白斑,疗效较满意。

关键词: 个体化

Abstract:

Objective To establish an individualized culture system for melanocytes, and to estimate its efficacy for the treatment of large-area vitiligo. Methods Hu16 medium was used for in vitro primary culture of melanocytes isolated from patients with stable segmental vitiligo. Doubling time (DOT), melanin content (M), melanin production (MP) and number of dendrites were examined to evaluate the biological activity of melanocytes. To obtain melanocytes with better biological activity, the components of Hu16 culture medium were adjusted. Ultra pulse CO2 laser was utilized to shave the vitiligous lesions and remove the epidermis followed by autologous transplantation. Follow-up was carried out. Results Melanocytes were obtained from 10 patients with stable segmental vitiligo and cultured. The melanocytes from 6 patients showed relatively short DOT, stable M and MP during the first and seventh passage, and were considered to be at initial or growth stage and applicable to transplantation. The remaining melanocytes from the other 4 patients had displayed long DOT, instable M, MP and dendrite quantity since the third passage; by adjusting the components of culture medium, these cells were induced into growth stage and finally applied to transplantation. A 12-month follow-up revealed that the repigmentation rate was higher than 90% in 7 patients, ranged between 70% and 80% in the remaining 3 patients, with the transplantation area being 116.8 ± 75.6 cm2. Conclusions The individualized culture system with adjusted components in culture medium yields melanocytes with satisfying biological activity, which are proved to be effective for the treatment of large-area, segmental and stable vitiligo.

Key words: individualization