[Isolation, culture and identification of adult epidermal stem cell].
OBJECTIVE
To explore and establish a new method of isolation, culture, and identification of adult epidermal stem cell in vitro for the provision of seed-cells in tissue engineering of skin.


METHODS
Epidermis was obtained by digesting human foreskin with protease and it was dissociated into single cells with trypsin and ethylenediaminetetraacetic acid (EDTA). These single epidermis cells were inoculated onto human collagen IV coated flasks and cultured at 37 degrees C in a humidified atmosphere containing 5% CO(2). The nonadherent cells were rinsed off 10-15 minutes after inoculation. The adherent cells were observed under phase contrast microscope and electron microscope, and they were identified with immunocytochemical methods. Cell cycles of the adherent cells were determined with flow cytometry.


RESULTS
With phase contrast microscope, the rapidly adherent cells were observed to form colonies 24 hours after inoculation. Immunocytochemistry showed the rapidly adherent cells were positive for beta1-integrin and keratin 19. Cell cycles showed that about 86.83% cells were in resting state/pre-DNA-synthetic gap (G0/G1 phase). Electron microscopy revealed that the rapidly adherent cells were immature.


CONCLUSIONS
This study shows that adult epidermal stem cells could be isolated and cultured in vitro successfully.
Xie
Xiao-fan
XF
Burns Department, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shanxi, China.
Jia
Chi-yu
CY
Fu
Xiao-bing
XB
Liu
Hu-xian
HX
D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D000078404:Epidermal Cells; D006801:Humans