Human Amnion-derived Mesenchymal Stem Cells (HAMSCs) Promote Osteogenesis And Angiogenic Differentiation Of Human Adipose-derived Stem Cells (HASCs)

Objective: Our previous studies have found that HAMSCs can promote proliferation,osteogenesis and angiogenic differentiation of HBMSCs.HASCs have the ability for multiple differentiation potential.HASCs are more available and abundant than HBMSCs.In this study,we investigated the effects of HAMSCs on proliferation,osteogenesis and angiogenic differentiation of HASCs.We aim to provide new seed cells for bone tissue engineering and provide theoretical and experimental basis for bone regeneration.Methods: We collected the amniotic membrane from the placentas of subscribers.We isolated and obtained HAMSCs.HAMSCs were characterized via its surface makers by flow cytometry.We choosed passaged3-6 for experiments.The Transwell co-culture system was used to investigated the effects of HAMSCs on HASCs.Proliferation levels were observed by flow cytometry.The effect of osteogenesis was investigated by alkaline phosphatase(ALP)activity substrate assay,Alizarin red S staining,and QPCR analysis of osteogenic marker expression.We used western blotting to find the potential mechanism of osteogenesis.We added the specific inhibitor of extracellular signaling-regulated kinase 1/2(Erk1/2)signaling such as U0126.We authenticated the effect and mechanism of osteogenesis in reverse via western blotting,ALP activity substrate assay,Alizarin red S staining,and QPCR analysis of osteogenic marker expression.Moreover,we used the same co-culture system and added U0126,enzyme-linked immunosorbent assay(ELISA)and human umbilical vein endothelial cells(HUVECs)tube formation were used to suggested the effect and mechanism of angiogenic differentiation.Results:1)HAMSCs are fibroblast like cells which are adhered to the wall of culture dish.Cell surface markers were identified by flow cytometry,which proved HAMSCs to be mesenchymal stem cells.2)HAMSCs have the effects on HASCs not only in proliferation but also in osteogenesis,and the activation of Erk1/2 signaling pathway has something to do with it.3)HAMSCs promotes angiogenesis of HASC,its mechanism is also related to the activation of Erk1/2 signaling pathway.Conclusion: HAMSCs promote proliferation,osteogenesis and angiogenic differentiation of HASCs.The findings suggest HAMSCs new and excellent seed cells for bone tissue engineering.