title: Use of synthetic serum-free medium for culture of human dermal fibroblasts to establish an experimental system similar to living dermis.
abstract: In this study, we sought to establish a defined experimental system for fibroblast growth similar to that of the living dermis. To this end, we evaluated the growth and biochemical characteristics of fibroblasts cultured with serum-free HFDM-1, a finely tuned synthetic medium for human fibroblast culture. Three culture conditions were used to grow fibroblasts obtained from primary culture: (1) culture with Dulbecco's modified Eagle medium (DMEM) plus 10 % fetal bovine serum (serum-supplemented DMEM), (2) culture with DMEM (serum-free DMEM), and (3) culture with HFDM-1 (HFDM-1), and fibroblast morphology, growth, collagen type I production, and lipid composition were analyzed. Fibroblasts grown in HFDM-1 maintained cell numbers at nearly 100 % from days 14 to 21 and produced more collagen type I than cells grown in serum-supplemented and serum-free DMEM. Arachidonic acid (20:4) and total polyunsaturated fatty acids were lower in cells grown in serum-free DMEM and HFDM-1 than in serum-supplemented DMEM. These results suggested that HFDM-1 recapitulated growth conditions in the dermis better than traditional, serum-supplemented DMEM. In addition, the controlled chemical composition of HFDM-1 eliminated a potential source of variability in cell culture conditions.
authors.affiliation: Division of Plastic Surgery, Kobe University Graduate School of Medicine, 7-5-2, Kusunoki-cho, Chuo-ku, Kobe, 650-0017, Japan, s8015@nms.ac.jp.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Ejiri
authors.forename: Tadashi
authors.initials: T
authors.lastname: Nomura
authors.forename: Masumi
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Chiaki
authors.initials: C
authors.lastname: Tatsumi
authors.forename: Midori
authors.initials: M
authors.lastname: Imai
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Sakakibara
authors.forename: Hiroto
authors.initials: H
authors.lastname: Terashi