PMID- 24137299
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20211021
IS  - 1792-0981 (Print)
IS  - 1792-1015 (Electronic)
IS  - 1792-0981 (Linking)
VI  - 6
IP  - 4
DP  - 2013 Oct
TI  - Optimization of the method for the culture of melanocyte precursors from hair 
      follicles and their activation by 1,25-dihydroxyvitamin D3.
PG  - 967-972
AB  - The melanocytes in vitiligo repigmentation are derived predominantly from 
      melanocyte precursors (MPs) present in the outer root sheath (ORS) of hair 
      follicles. The methods currently used for culturing MPs are unstable, and the 
      cultured cells have the capacity to produce melanin. These factors are 
      problematic when conducting in vitro studies to investigate the mechanism of 
      repigmentation. Although 1,25-dihydroxyvitamin D3 (VID) has been demonstrated to 
      be highly effective in the treatment of vitiligo in the clinic, its precise mode 
      of action has yet to be elucidated. In the present study, the method for the 
      culture of MPs from the ORS of hair follicles was optimized and the ability of 
      VID to activate MPs was investigated. The results suggested that the MPs cultured 
      using the optimized method mainly exhibited bipolar morphology. The cells 
      proliferated well and were negative for 3,4-dihydroxy-L-phenylalanine (DOPA) 
      staining. Transmission electron microscopy revealed that the cytoplasm of the MPs 
      contained numerous stage I and stage II melanosomes; however, stage III and IV 
      melanosomes were not observed. Following VID treatment, the MPs showed increased 
      dendritic morphology, the cells stained positive for DOPA and stage III and IV 
      melanosomes appeared in the cells. Western blotting revealed that 
      microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), 
      tyrosinase-related protein-1 (TRP-1) and TRP-2 were expressed in the MPs and that 
      VID increased the expression levels of MITF, TYR and TRP-1. However, the levels 
      of MITF, TYR and TRP-1 in the MPs prior to and following VID treatment were 
      significantly lower compared with those in cultured epidermal melanocytes, while 
      the levels of TRP-2 in these three groups were not significantly different. 
      Subsequent to VID treatment, the TYR activity in the MPs increased significantly, 
      as did the corresponding melanin levels. In conclusion, the present study 
      successfully optimized the method for MP culture. The MPs demonstrated no 
      significant TYR activity or melanin synthesis; therefore, the MP cultures 
      exhibited the features of MPs in vivo. In addition, VID significantly promoted 
      the differentiation of MPs.
FAU - Wang, Daguang
AU  - Wang D
AD  - Department of Dermatology, The First Affiliated Hospital of Nanjing Medical 
      University, Nanjing, Jiangsu 210029;
FAU - Xu, Xihui
AU  - Xu X
FAU - Ma, Huijun
AU  - Ma H
FAU - Yue, Xuezhuang
AU  - Yue X
FAU - Li, Chengrang
AU  - Li C
FAU - Zhu, Wenyuan
AU  - Zhu W
LA  - eng
PT  - Journal Article
DEP - 20130806
PL  - Greece
TA  - Exp Ther Med
JT  - Experimental and therapeutic medicine
JID - 101531947
PMC - PMC3797309
OTO - NOTNLM
OT  - 1,25 dihydroxyvitamin D3
OT  - activation
OT  - culture
OT  - melanocyte precursors
EDAT- 2013/10/19 06:00
MHDA- 2013/10/19 06:01
PMCR- 2013/08/06
CRDT- 2013/10/19 06:00
PHST- 2013/04/09 00:00 [received]
PHST- 2013/07/24 00:00 [accepted]
PHST- 2013/10/19 06:00 [entrez]
PHST- 2013/10/19 06:00 [pubmed]
PHST- 2013/10/19 06:01 [medline]
PHST- 2013/08/06 00:00 [pmc-release]
AID - etm-06-04-0967 [pii]
AID - 10.3892/etm.2013.1252 [doi]
PST - ppublish
SO  - Exp Ther Med. 2013 Oct;6(4):967-972. doi: 10.3892/etm.2013.1252. Epub 2013 Aug 6.