Objective: To evaluate the effects of low molecular weight heparin (LMWH) and heparin-binding epidermal growth factor (HB-EGF) on the biological function of human trophoblast in first trimester.
Methods: From Feb. 2011 to Nov. 2011, the trophoblast isolated from human first trimester chorionic villi was cultured in vitro. Based on variation of LMWH concentration, the trophoblast was classified into 0.025 U/ml group, 0.25 U/ml group, 2.5 U/ml group, 25 U/ml group and 250 U/ml group. In the mean time, based on treatment of heparin, the trophoblast was classified into LMWH group (0.25 U/ml), HB-EGF group (10 µg/L), combination group (LMWH at 0.25 U/ml + HB-EGF at 10 µg/L) and add with DMEM as control group. Cell proliferation was assessed by the methyl thiazolyl tetrazolium (MTT) test, which was showed with the mean absorbance as A value. Cell invasion was measured by transwell, which counted the number of cells migrated to the superficies inferia of filter membrane. Cell differentiation was assessed by the concentration of hCG secretion.
Results: Compared with control group, the trophoblast proliferation and invasion treated by LMWH at 0.025 U/ml did not show significant difference (P > 0.05). When treated by LWMH at 0.25 U/ml and 2.5 U/ml, trophoblast proliferation and invasion was increased significantly (P < 0.05). When LMWH at 25 U/ml and 250 U/ml, it could inhibit trophoblast proliferation and invasion (P < 0.05). When compared with A value of 0.44 ± 0.04 in control group, the increased A value were 0.51 ± 0.05 in LMWH group, 0.56 ± 0.04 in HB-EGF group and 0.69 ± 0.06 in combination group (P < 0.05). In the transwell test, the cell number were 511 ± 78 in LMWH group, 669 ± 67 in HB-EGF group and 872 ± 64 in combination group, which were significantly higher than 405 ± 67 in control group (P < 0.05), respectively. And the hCG concentration were (7143 ± 649) U/L in LMWH group, (11 762 ± 1059) U/L in HB-EGF group and (11 015 ± 1084) U/L in combination group, which showed statistical difference with (8182 ± 666) U/L in control group (P < 0.05).
Conclusion: LMWH could modulate trophoblast proliferation, invasion, and differentiation. HB-EGF is one of important factors involved in effects of LMWH on trophoblast function.