title: Synergistic effects of FGF-2 with insulin or IGF-I on the proliferation of human auricular chondrocytes.
abstract: Chondrocyte preparation with the safety and efficiency is the first step in cartilage regenerative medicine. To prepare a chondrocyte proliferation medium that does not contain fetal bovine serum (FBS) and that provides more than a 1000-fold increase in cell numbers within approximately 1 month, we attempted to use the medium containing 5% human serum (HS), but it exerted no more than twofold increase in 2 weeks. To compensate for the limited proliferation ability in HS, we investigated the combinational effects of 12 factors [i.e., fibroblast growth factor (FGF)-2, insulin-like growth factor (IGF)-I, insulin, bone morphogenetic protein-2, parathyroid hormone, growth hormone, dexamethasone, 1alpha25-dihydroxy vitamin D3, L-3,3',5'-triodothyronine, interleukine-1 receptor antagonist, 17beta-estradiol, and testosterone] on the proliferation of human auricular chondrocytes by analysis of variance in fractional factorial design. As a result, FGF-2, dexamethasone, insulin, and IGF-I possessed promotional effects on proliferation, while the combination of FGF-2 with insulin or IGF-I synergistically enhanced the proliferation. Actually, the chondrocytes increased 7.5-fold in number in 2 weeks in a medium containing 5% HS with 10 ng/ml FGF-2, while the cell number synergistically gained a 10-12-fold increase with 5 microg/ml insulin or 100 ng/ml IGF-I in the same period. The proliferation effects were more enhanced at a concentration of 100 ng/ml for FGF-2, and especially for the combination of 100 ng/ml FGF-2 and 5 microg/ml insulin (approximately 16-fold within 2 weeks). In the long-term culture with repeated passaging, this combination provided more than 10,000-fold within 8 weeks (i.e., passage 4). Thus, we concluded that such a combination of FGF-2 with insulin or IGF-I may be useful for promotion of auricular chondrocyte proliferation in a clinical application for cartilage regeneration.
authors.affiliation: Department of Fujisoft ABC [corrected] Cartilage & Bone Regeneration, Graduate School of Medicine [corrected] University of Tokyo, Hongo 7-3-1, Bunkyo-Ku, Tokyo 113-0033 [corrected] Japan.
authors.forename: Tsuguharu
authors.initials: T
authors.lastname: Takahashi
authors.forename: Toru
authors.initials: T
authors.lastname: Ogasawara
authors.forename: Junji
authors.initials: J
authors.lastname: Kishimoto
authors.forename: Guangyao
authors.initials: G
authors.lastname: Liu
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Asato
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakatsuka
authors.forename: Eijyu
authors.initials: E
authors.lastname: Uchinuma
authors.forename: Kozo
authors.initials: K
authors.lastname: Nakamura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kawaguchi
authors.forename: Ung-il
authors.initials: UI
authors.lastname: Chung
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Takato
authors.forename: Kazuto
authors.initials: K
authors.lastname: Hoshi
mesh_terms: D000293:Adolescent; D000704:Analysis of Variance; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D003470:Culture Media; D004357:Drug Synergism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D023822:Tissue Engineering / Q000379:methods
chemical_list: D003470:Culture Media; D007328:Insulin; D016222:Fibroblast Growth Factor 2; D007334:Insulin-Like Growth Factor I