Human keratinocytes are efficiently immortalized by a Rho kinase inhibitor

Sandra Chapman, Xuefeng Liu, Craig Meyers, Richard Schlegel, Alison A. McBride

Research output: Contribution to journalArticlepeer-review

252 Scopus citations


Primary human keratinocytes are useful for studying the pathogenesis of many different diseases of the cutaneous and mucosal epithelia. In addition, they can form organotypic tissue equivalents in culture that can be used as epidermal autografts for wound repair as well as for the delivery of gene therapy. However, primary keratinocytes have a finite lifespan in culture that limits their proliferative capacity and clinical use. Here, we report that treatment of primary keratinocytes (originating from 3 different anatomical sites) with Y-27632, a Rho kinase inhibitor, greatly increased their proliferative capacity and resulted in efficient immortalization without detectable cell crisis. More importantly, the immortalized cells displayed characteristics typical of primary keratinocytes; they had a normal karyotype and an intact DNA damage response and were able to differentiate into a stratified epithelium. This is the first example to our knowledge of a defined chemical compound mediating efficient cell immortalization, and this finding could have wide-ranging and profound investigational and medical applications.

Original languageEnglish (US)
Pages (from-to)2619-2626
Number of pages8
JournalJournal of Clinical Investigation
Issue number7
StatePublished - Jul 1 2010

All Science Journal Classification (ASJC) codes

  • General Medicine


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