title: Development of a chemically defined in vitro culture system to effectively stimulate the proliferation of adult human dermal fibroblasts.
abstract: 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 aHDFs. KDMEM supplemented with 5% (v/v) KSR, 12 ng/ml bFGF, 5 ng/ml EGF and 1 μg/ml hydrocortisone supported sufficient proliferation of aHDFs for 1 week. However, aHDF proliferation was decreased greatly after subculture. This problem could be overcome by culturing aHDFs in CDCM in culture plates coated with 10 μg/ml FN. Long-term culture of aHDFs was achieved using CDCM and FN-coated culture plates for 7 weeks. The optimized CDCS increased the proliferation of aHDFs significantly, without any increase in the senescence rate or alteration in morphology of aHDFs, despite long-term culture. In conclusion, we established a CDCS that improved proliferation of aHDFs while inhibiting cellular senescence. The CDCS will contribute to advances in various future research related to clinical skin regeneration.
authors: Department of Animal Life Science, Kangwon National University, Chuncheon, Korea.
authors: Min Seong
authors: MS
authors: Kim
authors: College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.
authors: Jung Im
authors: JI
authors: Yun
authors: Department of Marine Biomaterials and Aquaculture, Pukyong National University, Busan, Korea.
authors: Seung Pyo
authors: SP
authors: Gong
authors: Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea.
authors: Ji Yeon
authors: JY
authors: Ahn
authors: Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea.
authors: Jeong Mook
authors: JM
authors: Lim
authors: Department of Animal Life Science, Kangwon National University, Chuncheon, Korea.|Division of Animal Resource Science, Kangwon National University, Chuncheon, Korea.
authors: Young Han
authors: YH
authors: Song
authors: Department of Animal Life Science, Kangwon National University, Chuncheon, Korea.|Division of Animal Resource Science, Kangwon National University, Chuncheon, Korea.
authors: Kyu Hyun
authors: KH
authors: Park
authors: Department of Animal Life Science, Kangwon National University, Chuncheon, Korea.|Division of Applied Animal Science, Kangwon National University, Chuncheon, Korea.
authors: Seung Tae
authors: ST
authors: Lee
mesh_terms: D000328:Adult; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D016922:Cellular Senescence; D016895:Culture Media, Serum-Free / Q000737:chemistry*; D005347:Fibroblasts / Q000166:cytology*; D005353:Fibronectins / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D012867:Skin / Q000166:cytology*
chemical_list: D016895:Culture Media, Serum-Free; D005353:Fibronectins
keywords: adult human dermal fibroblasts; chemically defined medium; knockout serum replacement; proliferation; senescence