Isolating human dermal fibroblasts using serial explant culture

Fereshteh Nejaddehbashi, Vahid Bayati, Leila Mashali, Mahmoud Hashemitabar, Mohammadreza Abbaspour, Eskandar Moghimipour, Mahmoud Orazizadeh


Background: The purpose of this study was to introduce an applicable culture technique to isolate human dermal fibroblasts (HDFs); which could also contribute to research, clinical practices, as well as tissue engineering.
Methods: Samples from the human skin were dissected and cultured via serial explant technique. Subsequently, the isolated fibroblasts were assessed for their protein markers and genetic variations via immunofluorescence (IF) and karyotyping; respectively. Following the employment of this technique, a small piece of explant completely disappeared; while no dermis remained after 10 days.
Results: The quantity of HDFs harvested through this culture technique was reported at a normal level. The results of immunostaining also indicated that the isolated fibroblasts had expressed vimentin and fibronectin; whereas no cells had shown cytokeratin and epidermal marker. Moreover, karyotyping results for the fibroblasts isolated by the given technique revealed no chromosomal diversity after passage 20.
Conclusions: It was concluded that serial explant culture was an efficient technique for isolating HDFs from a small piece of skin in short-time periods; which could also preserve their normal morphology and molecular characteristics.