Human keratinocytes are efficiently  immortalized by a Rho kinase inhibitor.
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.
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland 20892, USA.
Sandra
S
Chapman
Xuefeng
X
Liu
Craig
C
Meyers
Richard
R
Schlegel
Alison A
AA
McBride
D000577:Amides; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D004848:Epithelium; D015316:Genetic Therapy; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism*; D011725:Pyridines; D012867:Skin; D014945:Wound Healing / Q000235:genetics; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors
D000577:Amides; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases