[Isolation, culture and identification of adult epidermal stem cell]

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.

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.

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.

This study shows that adult epidermal stem cells could be isolated and cultured in vitro successfully.