Spontaneous immortalization of human epidermal cells with naturally elevated telomerase

Miguel A. Rea, Lei Zhou, Qin Qin, Yann Barrandon, Karen W. Easley, Sarah F. Gungner, Marjorie A. Phillips, William S. Holland, Paul H. Gumerlock, David M Rocke, Robert H. Rice

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Abstract

This work explores spontaneous immortalization in keratinocytes, derived from two skin samples, that display naturally elevated telomerase activity. Serially passaged with 3T3 feeder layer support, the keratinocytes were examined for colony-forming ability, telomerase activity, telomere length, and finally gene expression using Affymetrix DNA microarrays. The cells initially exhibited normal karyotypes and low colony-forming efficiencies typical of normal epidermal cells, but after 40 passages (≈ 400 generations) colony-forming ability increased markedly, yielding immortalized lines exhibiting a small number of chromosomal aberrations and functionally normal p53. An improved protocol for analysis of microarray data permitted detection of 707 transcriptional changes accompanying immortalization including reduced p16 INK4A mRNA. Telomerase activity was clearly elevated in cells even at low passage from both samples, and telomerase catalytic subunit mRNA was greatly elevated in those with elevated colony-forming ability. The data raise the possibility of an unusual natural phenotype in which aberrant telomerase regulation extends keratinocyte lifespan until rare variants evade senescence. In addition to revealing a potential tumor-prone syndrome, the findings emphasize the desirability of carefully minimizing the degree or timing of elevated expression of telomerase used to immortalize cells for therapeutic purposes.

Original languageEnglish (US)
Pages (from-to)2507-2515
Number of pages9
JournalJournal of Investigative Dermatology
Volume126
Issue number11
DOIs
StatePublished - Nov 15 2006

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ASJC Scopus subject areas

  • Dermatology

Cite this

Rea, M. A., Zhou, L., Qin, Q., Barrandon, Y., Easley, K. W., Gungner, S. F., Phillips, M. A., Holland, W. S., Gumerlock, P. H., Rocke, D. M., & Rice, R. H. (2006). Spontaneous immortalization of human epidermal cells with naturally elevated telomerase. Journal of Investigative Dermatology, 126(11), 2507-2515. https://doi.org/10.1038/sj.jid.5700424