Synergistic effects of FGF-2 with insulin or IGF-I on the proliferation of human auricular chondrocytes
Synergistic effects of FGF-2 with insulin or IGF-I on the proliferation of human auricular chondrocytes
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DOI：
10.3727/000000005783982675
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发表时间：
2005-01-01
影响因子：
3.3
通讯作者：
Hoshi, K
中科院分区：
文献类型：
--
作者：
Takahashi, T;Ogasawara, T;Hoshi, K
安全、高效的软骨细胞制备是软骨再生医学的第一步。为了制备不含胎牛血清（FBS）并在约1个月内提供超过1000倍的细胞数增加的软骨细胞增殖培养基，我们尝试使用含有5%人血清（HS）的培养基，但其在2周内产生不超过2倍的增加。为了补偿HS中有限的增殖能力，我们研究了12个因素的组合效应[即，成纤维细胞生长因子（FGF）-2、胰岛素样生长因子（IGF）-I、胰岛素、骨形态发生蛋白-2、甲状旁腺激素、生长激素、地塞米松、1 α 25-二羟基维生素D-3、L-3，3 '，5'-三碘甲腺原氨酸、白细胞介素-1受体拮抗剂、17 β-雌二醇和睾酮]对人耳廓软骨细胞增殖的影响。结果，FGF-2、地塞米松、胰岛素和IGF-I对增殖具有促进作用，而FGF-2与胰岛素或IGF-I的组合协同增强增殖。实际上，在含有5%HS和10 ng/ml FGF-2的培养基中，软骨细胞的数量在2周内增加了7.5倍，而在同一时期，细胞数量在5 μ g/ml胰岛素或100 ng/ml IGF-1的情况下协同获得了10-12倍的增加。对于FGF-2，增殖作用在100 ng/ml的浓度下更增强，特别是对于100 ng/ml FGF-2和5 μ g/ml胰岛素的组合（2周内约16倍）。在重复传代的长期培养中，该组合在8周内提供超过10，000倍（即，第4段）。因此，我们得出结论，FGF-2与胰岛素或IGF-I的这种组合在软骨再生的临床应用中可用于促进耳廓软骨细胞增殖。
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 I 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, 1 alpha 25-dihydroxy vitamin D-3, L-3,3',5'-triodothyronine, interleukine-1 receptor antagonist, 17 beta-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 mu g/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 mu g/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.