[Effect of three growth factors on proliferation and cell phenotype of human fetal meniscal cells]

Abstract

Objective: 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.

Methods: 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.

Results: 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).

Conclusion: 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.