PIG3V, an immortalized human vitiligo melanocyte cell line, expresses dilated endoplasmic reticulum.
Vitiligo is an enigmatic pigmentary disorder of the skin. Factors potentially involved in the progressive loss of melanocytes from the basal layer of the epidermis include genetically determined aberrancies of the vitiligo melanocyte. It follows that analysis of melanocytes cultured from vitiligo donors can contribute to a further understanding of the etiopathomechanism. A setback for vitiligo research has been the limited availability of vitiligo-derived melanocytes. To overcome this limitation, we have generated a vitiligo melanocyte cell line according to a protocol established previously for the immortalization of normal human melanocytes. Vitiligo melanocytes Ma9308P4 were transfected with HPV16 E6 and E7 genes using the retroviral construct LXSN16E6E7. Successful transformants were selected using geneticin and subsequently cloned to ensure genetic homogeneity. The resulting cell line PIG3V has undergone more than 100 cell population doublings since its establishment as a confluent primary culture, whereas untransfected melanocytes derived from adult skin senesce after a maximum of 50 population doublings. Cells immortalized by this transfection procedure retain lineage-specific characteristics and proliferate significantly faster than parental cells. In this study, the phenotype of PIG3V resembled melanocytes rather than melanoma cells in culture. Tyrosinase was processed properly and melanosomes remained pigmented. Importantly, ultrastructural characterization of PIG3V cells revealed dilated endoplasmic reticulum profiles characteristic of vitiligo melanocytes. An explanation for this dilation may be found in the retention of proteins with molecular weight of 37.5. 47.5, and 56.5 kDa, as determined by gel electrophoresis of microsomal proteins isolated from radiolabeled cells.
Department of Dermatology, Amsterdam University, The Netherlands.
I C
IC
Le Poole
R E
RE
Boissy
R
R
Sarangarajan
J
J
Chen
J J
JJ
Forristal
P
P
Sheth
W
W
Westerhof
G
G
Babcock
P K
PK
Das
C B
CB
Saelinger
D000328:Adult; D002461:Cell Line, Transformed; D002999:Clone Cells; D018870:Endoplasmic Reticulum, Rough; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008544:Melanocytes / Q000166:cytology*; D008854:Microscopy, Electron / Q000379:methods; D009856:Oncogene Proteins, Viral / Q000235:genetics; D050725:Papillomavirus E7 Proteins; D010858:Pigmentation; D012097:Repressor Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase; D014820:Vitiligo / Q000473:pathology*
C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D019098:Telomerase