Effect of SDF-1/HGF fusion protein-mediated BMSCs on proliferation and chemotaxis of HSCs

Abstract:

Objective：To investigate the effect of SDF-1/HGF fusion protein-mediated BMSCs on proliferation and chemotaxis of HSCs，then to rebuild liver tissue by combined transplantation. Methods：BMSCs and HSCs derived from SD rat at the age of 3-4 weeks were cultured and identified. After stably serial subcultivation，recombinated adenovirus pAD-SDF-1-（GlySer）3-HGF-IRES-GFP infected BMSCs，and the efficiency of infection was detected after infection for 1，3，7，14 d by fluorescence microscope. Meanwhile，the expressions of SDF-1 and HGF were assayed by ELISA. HSCs were planted in 24 well plates containing BMSCs cytotrophoblast. They were classified as HSCs plus cell culture fluid（group A1），HSCs plus BMSCs（group B1），HSCs plus SDF-1/HGF fusion protein-mediated BMSCs（group C1）. The proliferation index of HSCs after coculture for 1，3，7，14 d were investigated. In order to exam the immigration index of HSCs influenced by SDF-1/HGF fusion protein-mediated BMSCs，three experimental groups including HSCs plus cell culture fluid（group A2），HSCs plus BMSCs（group B2），HSCs plus SDF-1/HGF fusion protein-me－diated BMSCs（group C2） through transwell system were set. Results：After BMSCs being infected by recombinated adenovirus pAD-SDF-1-（GlySer）3-HGF-IRES-GFP for1，3，7，14 d，the efficiencies of infection were （25.28±3.72）%，82.22±3.58）%，（64.85±4.48）%，（28.35±2.73）%，respectively. And the efficiency of infection was the highest after infection for 3 d（com－pared with infection for 1，7，14 d，F=718.05，P＝0.000；F=66.80，P＝0.000；F=642.78，P＝0.000）. At the same time，after infection for 1，3，7，14 d，the expressions of SDF-1 were （1.02±0.15） μg/ml，（4.51±0.52） μg/ml，（2.13±0.19） μg/ml，（1.22±0.21） μg/ml，and the expressions of HGF were （1.38±0.32） μg/ml，（5.01±0.43） μg/ml，（3.10±0.26） μg/ml，（1.62±0.27） μg/ml. According to our data，after infection for 3 d，the expressions of SDF-1 and HGF were significantly increased compared with those after infection for 1，7，14 d（P＝0.000）. After coculture for 3，7，14 d，the proliferation of HSCs were significantly increased in group B1 and group C1（P＝0.000）compared with those of group A1. And the amplification of HSCs was obviously stronger in group C1 than in group B1（P＝0.000）. In transwell test，the immigration indexes of HSCs were prominently increased in group B2 and group C2 than in group A2. Meanwhile，the immigration index of HSCs in group C2 was remarkably higher than that in group B2. Conclusion：SDF-1/HGF fusion protein-mediated BMSCs might have an key role in promoting proliferation and chemotaxis of HSCs.