Growth of human melanocytes from suction blister of localized vitiligo patient and from foreskins of newborns and adults by modified culture medium.
We report on the culturing of melanocytes from suction blisters from the uninvolved skin of localized (focal and segmental) vitiligo patients and from the foreskins of newborns and adults, over a long period of time using a modified culture medium composed of F-12 medium supplemented with insulin (5 mg/mL), cholera toxin (40 ng/mL), transferrin (5 mg/mL), hydrocortisone (1 mM), epithelial growth factor (20 ng/mL), endothelial cell growth supplement (ECGS) (15 mg/mL), retinol (1 x 10(-7) M), 12-o-tetradecanoyl-phorbol-13 acetates (TPA) (85 nM), and 1% fetal calf serum (FCS). This method may be used in in vitro studies on normal human melanocytes and for study of the difference between melanocytes of normal individuals and of those with vitiligo. The ability to culture melanocytes from localized vitiligo, and the inability to grow those of generalized vitiligo with this system reconfirms the difference in the pathogenesis of various types of vitiligo. Thus, culturing may be used for differentiating localized vitiligo from generalized vitiligo and may be used to guide the mode of treatment when vitiligo has just started to develop and only a few depigmented patches have appeared. The results of studies on seeding density effect and serial Dopa reaction on the melanocytes of normal individuals reveal that the use of tyrosinase activity as the assessment parameter requires the use of melanocytes cultured for less than five months at a seeding density of less than 4 x 10(4) cells/cm2. Enlarged and heavily pigmented cells, senescent cells, were observed after very long-term culture (one year and four months).(ABSTRACT TRUNCATED AT 250 WORDS)
Jee
S H
SH
Department of Dermatology, National Taiwan University, Taipei, R.O.C.
Kuo
Y F
YF
Wu
Y C
YC
Chang
C H
CH
Kuo
W L
WL
Lü
Y C
YC
D000293:Adolescent; D000328:Adult; D002452:Cell Count; D002478:Cells, Cultured; D002648:Child; D003470:Culture Media; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008544:Melanocytes / Q000473:pathology* / Q000502:physiology; D008875:Middle Aged; D014820:Vitiligo / Q000473:pathology* / Q000503:physiopathology
D003470:Culture Media