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Establishing melanocyte cultures in a serum-free system for transplantation in vitiligo patients.

Rafał Czajkowski, Waldemar Placek, Tomasz Drewa, Dorota Olszewska-Słonina, Jan Sir, Bogna Kowaliszyn, Marcin Weiss

Med Sci Monit 2006; 12(2): CR63-69

ID: 445238

Available online: 2006-02-01

Published: 2006-02-01

Background: The aim of the study was to establish melanocyte cultures obtainedfrom the epidermis of vitiligo patients for future treatment and to estimate the influence of selectedfactors on the formation of suction blisters and the results of culture. Material/Methods: Twenty patientswith generalized vitiligo were qualified to the study. The melanocyte donor site was the arm or forearmwhere, using an electric vacuum suction machine and a plastic plate, suction blisters were formed. Theremoved roofs of the blisters were used as a melanocyte source for culture establishment. After an enzymaticbath, the cell suspension was transferred to culture flasks containing melanocyte growth medium (MGMM2). The melanocytes in culture were identified by immunocytochemistry. Results: Suction blisters wereformed in all patients. No melanocyte cultures were established from patients whose blisters appearedover a period 58 minutes. In addition, a suitable quantity of melanocytes was not obtained for eightother patients (with skin phototype II and/or without previous PUVA therapy). Conclusions: The epidermisof the arm and forearm is a good donor site for the establishment of melanocyte cultures. The effectivenessof culture depends mainly on the time of the formation of suction blisters, the phototype of the skin,and previous PUVA therapy.

Keywords: Arm, Adult, Cell Culture Techniques - methods, Cell Separation, Culture Media, Serum-Free, Epidermis - pathology, Melanocytes - transplantation, PUVA Therapy, Phenotype, Suction, Transplantation, Autologous, Vitiligo - surgery