Efficient procurement of epithelial stem cells from human tissue specimens using a Rho-associated protein kinase inhibitor Y-27632.
The efficient culture of stem cells from epithelial tissues such as skin and corneas is important for both experimental studies and clinical applications of tissue engineering. We now demonstrate that treatment of human-skin-derived keratinocytes with a Rho-associated protein kinase inhibitor Y-27632 for the initial 6 days of primary culture can increase the number of keratinocytes that possess stem cell properties to form colonies during in vitro culture of freshly isolated cells and subsequent passage (50-fold). Further, we show that Y-27632 treatment can increase the total number of prostate epithelial cells derived from human prostate specimens. Therefore, the use of Y-27632 during primary cultures offers a simple and effective way to prepare a large number of epithelial stem cells from various human epithelial tissues.
Terunuma
Atsushi
A
Dermatology Branch, Center for Cancer Research, National Cancer Institute , National Institutes of Health, Bethesda, MD, USA. atsushi@mail.nih.gov
Limgala
Renuka Pudi
RP
Park
Christine J
CJ
Choudhary
Isha
I
Vogel
Jonathan C
JC
D016475:3T3 Cells; D000577:Amides / Q000494:pharmacology*; D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D018920:Coculture Techniques; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D052816:Foreskin / Q000166:cytology; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D008297:Male; D051379:Mice; D011467:Prostate / Q000166:cytology; D011725:Pyridines / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D023822:Tissue Engineering; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
D000577:Amides; D004791:Enzyme Inhibitors; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases