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Short exposure to collagenase and coculture with mouse embryonic pancreas improve human dermal fibroblast culture
Author(s) -
Pandamooz Sareh,
Hadipour Abbas,
AkhavanNiaki Haleh,
Pourghasem Mohsen,
Abedian Zeinab,
Ardekani Ali Motevallizadeh,
Golpour Monireh,
Hassan Zuhair Mohammad,
Mostafazadeh Amrollah
Publication year - 2012
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1020
Subject(s) - collagenase , fibroblast , dispase , connective tissue , biology , embryonic stem cell , foreskin , dermal fibroblast , explant culture , cell culture , in vitro , microbiology and biotechnology , andrology , biochemistry , enzyme , medicine , genetics , gene
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 mm 3 ) 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.