title: Short exposure to collagenase and coculture with mouse embryonic pancreas improve human dermal fibroblast culture.
abstract: The dermal fibroblast as a major component of connective tissue has attracted much attention in the past few years, and application of these very fast growing cells in several fields has been intensively studied. Isolating dermal fibroblasts is an appropriate way to expand these fast growing cells in vitro. Although using a dissociated fibroblast culture method is more convenient than skin explant culture, its enzymatic digestion is critical because a large number of cells can be lost over prolonged exposure to collagenase. This study was performed to increase the number of viable cells after digestion of fresh human foreskin of donors aged from 1 to 3 months with collagenase and also by to design a coculture system for resuscitation of the injured fibroblast. Our results demonstrate that we can maximize cell yield while maintaining cell viability by cutting the specimens into very small pieces (1-2 mm³) after removing the epidermal layer with dispase II and also by collecting released cells every 20 Min subsequent to digesting the dermal layer with collagenase. Moreover, our data strongly indicate that coculturing of isolated fibroblasts with embryonic pancreas explants can enhance the rate of proliferation in cultured fibroblasts.
authors.affiliation: Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran.
authors.forename: Sareh
authors.initials: S
authors.lastname: Pandamooz
authors.forename: Abbas
authors.initials: A
authors.lastname: Hadipour
authors.forename: Haleh
authors.initials: H
authors.lastname: Akhavan-Niaki
authors.forename: Mohsen
authors.initials: M
authors.lastname: Pourghasem
authors.forename: Zeinab
authors.initials: Z
authors.lastname: Abedian
authors.forename: Ali Motevallizadeh
authors.initials: AM
authors.lastname: Ardekani
authors.forename: Monireh
authors.initials: M
authors.lastname: Golpour
authors.forename: Zuhair Mohammad
authors.initials: ZM
authors.lastname: Hassan
authors.forename: Amrollah
authors.initials: A
authors.lastname: Mostafazadeh
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival; D018920:Coculture Techniques; D017364:Collagenases / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology*; D052816:Foreskin / Q000166:cytology; D006801:Humans; D008297:Male; D051379:Mice; D010179:Pancreas / Q000166:cytology*; D013997:Time Factors
chemical_list: D017364:Collagenases