Generation of a human melanocyte cell line by introduction of HPV16 E6 and E7 genes

I. Caroline Le Poole*, Frank M. Van den Berg, René M J G J Van den Wijngaard, Denise A. Galloway, Peter J. Van Amstel, Anita A M Buffing, Henk L. Smits, Wiete Westerhof, Pranab K. Das

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

53 Scopus citations

Abstract

Availability of a standard human melanocyte cell line with unlimited growth potential and otherwise normal melanocytic properties will greatly facilitate research in melanocyte biology and in vitro studies on the etiology of pigmentary disorders and melanoma. Using a retroviral vector, E6 and E7 open reading frames of human papilloma virus type 16 (HPV 16) have been introduced into cultured normal human melanocytes. Cells selected by increased resistance to geneticin conveyed by the vector and expressing E6E7 mRNA have been cloned to ensure genetic homogeneity. Since their establishment as primary cells, cloned PIG1 cells have undergone more than twice the amount of population doublings of senescent parental cells. Moreover, in passage numbers when parental cells had become senescent, proliferation of clonal cells was retained at levels exceeding those of normal human melanocytes in third passage by 100%. Further characterization has revealed that the cells remain dependent on tetradecanoyl phorbol 13- acetate (TPA) for growth and do not proliferate in soft agar nor form tumors in nude mice. The antigenic profile of the cells was slightly altered as compared to parental cells, but was incomparable to that of M14 melanoma cells. Importantly, PIG1 cells contain more melanin pigment than parental cells.

Original languageEnglish (US)
Pages (from-to)42-49
Number of pages8
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume33
Issue number1
DOIs
StatePublished - Jan 1 1997

Keywords

  • HPV16E6E7
  • cell line
  • immortalization
  • pigmentation
  • proliferation

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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