ABSTRACT
Objective To investigate the protective function of paracrine hepatocyte growth factor (HGF) derived from human placental mesenchymal stem cells of fetal origin (hfPMSCs) cultured in serum-free medium against endothelial cell injury induced by lipopolysaccharide (LPS). Methods The hfPMSCs were cultured in serum-free medium and surface antigen CD73, CD90, CD105, CD14, CD34, CD45 and HLA-DR were analyzed by flow cytometry. Using TranswellTM co-culture system [human pulmonary microvascular endothelial cell (HPMECs) were added into the upper chambers of TranswellTM inserts, and hfPMSCs were added into the lower chambers of TranswellTM inserts], the influence of hfPMSCs paracrine HGF on the permeability of HPMECs in LPS condition was identified. Then four different co-culture conditions were used as follows: 100 ng/mL LPS treatment group; hfPMSCs co-culture group; HGF neutralization group; HPMECs normal control. After 100 µg FITC-dextran was added into the upper chambers of TranswellTM inserts, the effect of stain permeability was detected by fluorescence microplate reader. The expression of VE-cadherin, caveolin-1 and cleaved caspase-3, cleaved PARP-1 in HPMECs were measured by Western blot analysis. Results The hfPMSCs showed the classic morphology of mesenchymal stem cells and expressed the surface markers CD73, CD90 and CD105, but did not express CD14, CD34, CD45 and HLA-DR. Compared with LPS treatment group, the co-culture with hfPMSCs dramatically inhibited the permeability of HPMECs, significantly up-regulated the expression of VE-cadherin, and reduced the expression of caveolin-1, cleaved caspase-3, cleaved PARP1. In contrast, neutralizing HGF with anti-HGF antibody reversed the above effects of hfPMSCs-HPMECs co-culture. Conclusion Owing to paracrine HGF, hfPMSCs possess the capability to availably inhibit the permeability of HPMECs induced by LPS.
