title: Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin.
abstract: Currently, culture and growth keratinocytes are important stages in achieving a reliable and reproducible skin tissue. In the present study, two different methods, enzymatic and explant methods, for keratinocytes isolation from human foreskin were compared. Foreskins were cut into 2-3 mm pieces and placed in trypsin at 4°C overnight for separation of the epidermis from the dermis. Subsequently, these samples were divided into two groups: i) Keratinocytes separated from the epidermis by trypsin and ii) by the explant method. These keratinocytes were divided into two groups: i) With no feeder layer and ii) onto a type I collagen scaffold. The cells were evaluated using immunocytochemistry and 4',6-diamidine-2'-phenylindole dihydrochloride (DAPI) staining. In the enzymatic treatment, after 7-10 days no attached cells were found in the cell culture dishes. In the explant method, keratinocytes were separated after ~24 h, attached rapidly and formed big colonies into a collagen scaffold. In the absence of a feeder layer, small colonies were developed with rapid loss of proliferation within 2-3 days. Keratinocytes showed positive immunoreactivity for the pan-cytokeratin marker and keratinocytes' nuclei were clearly observed. This method could be applied and developed as a component of skin substitutes to treat burns and wounds and also in laboratory testing.
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran ; Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Orazizadeh
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran ; Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Hashemitabar
authors.affiliation: Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran.
authors.forename: Somayeh
authors.initials: S
authors.lastname: Bahramzadeh
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran.
authors.forename: Freshteh Nejad
authors.initials: FN
authors.lastname: Dehbashi
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-15794, Iran.
authors.forename: Sadegh
authors.initials: S
authors.lastname: Saremy
keywords: enzymatic method; explant method; human foreskin; keratinocytes