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.

Starved human fibroblasts secrete acidic proteins inducing post re‐feeding proliferation and in vitro cell migration: A potential tool for wound healing

“…Based on the method we described earlier (Pandamooz et al., ), fibroblasts were isolated from fresh foreskin prepared from 15 children aged 1.5–3 months (mean ± SD; 1.62 ± 0.62) who underwent routine circumcision from January 2012 to June 2012 at Amirkola Children's Hospital, in Babol, Iran.…”

Fabrication of PEO/chitosan/PCL/olive oil nanofibrous scaffolds for wound dressing applications

“…The cells were treated by culturing in DMEM high glucose (PAAE15-883) supplemented with 10 % fetal bovine serum (FBS, PAAA15-15), 1 % antibiotics penicillin and streptomycine (PAAP11-010) under standard conditions of 5 % CO 2 at 37 o C. The culture medium was changed every 3 days until passage 3 [31].…”

Antioxidant and Apoptotic Effects of an Aqueous Extract of Urtica dioica on the MCF-7 Human Breast Cancer Cell Line

“…The cells were cultured in RPMI-1640 medium (PAA, Austria) supplemented with 10% fetal bovine serum and 1% antibiotics (penicillin/ streptomycin (Invitrogen) in a humidified atmosphere containing 5%CO 2 and 95% air, at 37°C. Primary human Fibroblasts were isolated from fresh foreskin isolated from children aged between 1.5-30 months who underwent routine circumcision as previously described (Pandamooz et al, 2012). Briefly 1-2 mm 3 pieces of foreskin were incubated at 37°C for 2 hrs in a 15 ml falcon tube in the presence of 0.5% dispase II (Sigma-Aldrich) then digested with 0.1% crude collagenase (Sigma-Aldrich, C2674).…”