A modified method for purifying amelanotic melanocytes from human hair follicles
| العنوان: | A modified method for purifying amelanotic melanocytes from human hair follicles |
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| المؤلفون: | Wen-Yuan Zhu, Da-Guang Wang, Hui-Jun Ma, Cheng-Rang Li, Xue-Zhuang Yue |
| المصدر: | The Journal of Dermatology. 33:239-248 |
| بيانات النشر: | Wiley, 2006. |
| سنة النشر: | 2006 |
| مصطلحات موضوعية: | Adult, Male, Tyrosinase, Cell Culture Techniques, Cell Separation, Dermatology, General Medicine, Biology, Outer root sheath, Hair follicle, Molecular biology, Trypsinization, medicine.anatomical_structure, Dermis, Melanoblast, Immunology, medicine, Humans, Melanocytes, Female, Epidermis, Hair Follicle, Melanosome |
| الوصف: | We describe a modified method for establishing long-term pure cultures of amelanotic melanocytes (AMMC) derived from human hair follicles. Normal human corpse scalp (just after death, 1 h) was transected 1 mm below the epidermis, and hair follicles in the remaining dermis were isolated by a two-step enzyme treatment. Hair follicle cell suspensions were prepared by 0.50% trypsin treatment for 30 min and cultured in an optimized melanoblast proliferation nature mitogen medium. Cells attached to the substratum were mostly amelanotic melanocytic in character with small, bipolar shapes in the early stage; only a few keratinocytes and rare fibroblasts were observed. Keratinocytes were easily removed by differential trypsinization. After the third passage, the proliferating cells were all amelanotic melanocytes as confirmed by immunostaining with polyclonal antibodies to alphaPEP7h, which recognized the tyrosinase protein located on melanosomes and NKI/beteb, which is a pre-melanosomal antigen against synthetic peptides corresponding to the carboxyl termini of human melanosomal protein GP100. Cultured AMMC were highly positive to L-dopa reactivity after the addition of IBMX to the culture medium for 7 days. Many stage I and II melanosomes and occasional stage III melanosomes without stage IV melanosomes were found in the cytoplasm by transmission electron microscope. This modified technique is potentially more suitable for cultivating amelanotic melanocytes. The availability of pure cultures of hair-follicle amelanotic melanocytes will facilitate investigations of the roles of those cells in migration and differentiation during treatment of vitiligo. |
| تدمد: | 1346-8138 0385-2407 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::6bc48eadc7823f78d44ec7ef894c870e https://doi.org/10.1111/j.1346-8138.2006.00059.x |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....6bc48eadc7823f78d44ec7ef894c870e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13468138 03852407 |
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