Premium
Development of a chemically defined in vitro culture system to effectively stimulate the proliferation of adult human dermal fibroblasts
Author(s) -
Kim Min Seong,
Yun Jung Im,
Gong Seung Pyo,
Ahn Ji Yeon,
Lim Jeong Mook,
Song Young Han,
Park Kyu Hyun,
Lee Seung Tae
Publication year - 2015
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/exd.12695
Subject(s) - subculture (biology) , in vitro , cell growth , senescence , microbiology and biotechnology , cell culture , regeneration (biology) , tissue culture , biology , andrology , chemistry , medicine , biochemistry , genetics
Despite the fact that dermal fibroblasts are a practical model for research related to cell physiology and cell therapy, an in vitro culture system excluding serum, which complicates standardization and specificity and induces variability and unwanted effects, does not exist. We tried to establish a CDCS that supports effective proliferation of aHDF s. KDMEM supplemented with 5% (v/v) KSR, 12 ng/ml bFGF , 5 ng/ml EGF and 1 μ g/ml hydrocortisone supported sufficient proliferation of aHDF s for 1 week. However, aHDF proliferation was decreased greatly after subculture. This problem could be overcome by culturing aHDF s in CDCM in culture plates coated with 10 μ g/ml FN. Long‐term culture of aHDF s was achieved using CDCM and FN‐coated culture plates for 7 weeks. The optimized CDCS increased the proliferation of aHDF s significantly, without any increase in the senescence rate or alteration in morphology of aHDF s, despite long‐term culture. In conclusion, we established a CDCS that improved proliferation of aHDF s while inhibiting cellular senescence. The CDCS will contribute to advances in various future research related to clinical skin regeneration.