Ye Chuan, Deng Zhansheng, Li Baojun
Department of Orthopedics, Xiangya Hospital of Central South University, Changsha Hunan 410008, PR China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2007 Oct;21(10):1137-41.
To investigate the effects of the insulin-like growth factor 1 (IGF-1), the transforming growth factor beta1 (TGF-beta1), and the basic fibroblast growth factor (bFGF) on proliferation and cell phenotype of the human fetal meniscal cells, and to find out the best combination and concentration of the growth factors for the meniscus tissue engineering.
The fetus came from the healthy woman accidental abortion and the procedure had got her approval. The human fetal meniscal fibrochondrocytes were cultured in vitro. The cell phenotype was identified by the collagen type II immunohistochemistry and Aggrecan immunofluorescence. In the growth factor groups, the 3rd passage meniscal cells synchronized by the serum starvation method and were mixed with IGF-1 (1, 10, 50, 100 microg/L), TGF-beta1 (0.1, 1.0, 5.0, 10.0, 50.0 microg/L), and bFGF (5, 10, 50, 100, 200 microg/L), respectively, and in the combination groups, the combinations of bFGF and TGF-beta1, bFGF and IGF-1, TGF-beta1 and IGF-1 were established at their optimal effect concentrations. The control group was also established for comparison. The dose-response relationship was studied at 48 h and 72 h by the MTT colorimetric method.
The 3rd passage meniscal cells could express collagen type II and Aggrecan before and after the addition of the three growth factors. The proliferating effects of the growth factors (IGF-1 50 microg/L, TGF-beta1 5 microg/L, bFGF 50 microg/L) on the 3rd passage cells at 48 h and 72 h were significantly better in the growth factor groups than in the control group (P < 0.05), and the combination groups of bFGF 50 microg/L and IGF-1 50 microg/L, IGF-1 50 microg/L and TGF-beta1 5 microg/L showed a significantly higher proliferating effect than that in the single growth factor group (P < 0.05). bFGF 50 microg/L and TGF-beta1 5 microg/L had no synergetic effect (P > 0.05).
IGF-1, TGF-beta1 and bFGF can promote the proliferation of the human fetal meniscal cells, respectively, and the combinations of bFGF and IGF-1, IGF-1 and TGF-beta1 at their optimal concentrations can have better proliferating effects than the single growth factor. They can be used for the in vitro amplification of the meniscal seed cells.
研究胰岛素样生长因子1（IGF-1）、转化生长因子β1（TGF-β1）和碱性成纤维细胞生长因子（bFGF）对人胎儿半月板细胞增殖及细胞表型的影响，找出用于半月板组织工程的生长因子最佳组合及浓度。
胎儿来自健康女性的意外流产且该操作已获其同意。体外培养人胎儿半月板纤维软骨细胞。通过Ⅱ型胶原免疫组织化学和聚集蛋白聚糖免疫荧光鉴定细胞表型。在生长因子组中，将经血清饥饿法同步化的第3代半月板细胞分别与IGF-1（1、10、50、100μg/L）、TGF-β1（0.1、1.0、5.0、10.0、50.0μg/L）和bFGF（5、10、50、100、200μg/L）混合，在联合组中，将bFGF与TGF-β1、bFGF与IGF-1、TGF-β1与IGF-1按其最佳效应浓度进行组合。同时设立对照组进行比较。采用MTT比色法于48小时和72小时研究剂量反应关系。
添加三种生长因子前后，第3代半月板细胞均可表达Ⅱ型胶原和聚集蛋白聚糖。生长因子组中，48小时和72小时时生长因子（IGF-1 50μg/L、TGF-β1 5μg/L、bFGF 50μg/L）对第3代细胞的增殖作用明显优于对照组（P<0.05），bFGF 50μg/L与IGF-1 50μg/L、IGF-1 50μg/L与TGF-β1 5μg/L的联合组增殖作用明显高于单一生长因子组（P<0.05）。bFGF 50μg/L与TGF-β1 5μg/L无协同作用（P>0.05）。
IGF-1、TGF-β1和bFGF均可分别促进人胎儿半月板细胞增殖，bFGF与IGF-1、IGF-1与TGF-β1在最佳浓度下的联合作用比单一生长因子具有更好的增殖效果。它们可用于半月板种子细胞的体外扩增。
