Synergistic effects of FGF-2 with insulin or IGF-I on the proliferation of human auricular chondrocytes.
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.
Takahashi
Tsuguharu
T
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.
Ogasawara
Toru
T
Kishimoto
Junji
J
Liu
Guangyao
G
Asato
Hirotaka
H
Nakatsuka
Takashi
T
Uchinuma
Eijyu
E
Nakamura
Kozo
K
Kawaguchi
Hiroshi
H
Chung
Ung-il
UI
Takato
Tsuyoshi
T
Hoshi
Kazuto
K
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
D003470:Culture Media; D007328:Insulin; D016222:Fibroblast Growth Factor 2; D007334:Insulin-Like Growth Factor I