Placental Mesenchymal Stem Cells In Vitro Isolation, Cultivation And Differentiation Of Osteogenic Cells And Nerve Cells

Objective:Human placenta-derived mesenchymal stem cells(hPMSCs)were isolated and purified to study the biological characteristics,then established the co-culture system between hPMSCs and osteoblasts or neural cells in vitro and study the influence of osteoblasts or neural cells on the differentiation of hPMSCs into osteoblasts or neural cells in the co-culture system.Methods:Mesenchymal stem cells (MSCs) were isolated and purified from human term placenta using collagenase digestion and identified by cell surface marker, cell growth curve,cell ultrastructure and osteogenesis; Osteoblasts or neural cells were incubated in Transwell double-deck culture dish in the co-culture group. hPMSCs were incubated in the upper layer in high glucose DMEM medium with 10% FBS and then the induced cells were identified by Alkaline phosphatase staining or NSE.hPMSCs were incubated in both layers in the control group.Result:1.Under the same conditions, the successful culture rate was 100% in group D, and 20%,60% and 80% in group A, B and C, respectively. The time of stretching and the time of primary culture with Hespan were shorter in the group D than that in other group.2.MSCs isolated from human term placenta were fibrous long spindle-like adherent cells and had strong proliferative ability;They were Strongly positive for CD44,CD29, but negative for CD34,CD106。3.they also have the potential to differentiate into fat cells and the oil red O staining were positive. Conclusion:A large number of adherent cells were visible as fibrous growth after 14 days; CD44,CD29 were positive strongly, CD34,CD45 were negtive; hPMSCs were differentiated into osteoblasts; Alkaline phosphatase staining of hPMSCs in the co-culture group was positive but in the control group was negative,the cells co-cultured with neural cells became slender and had the filament, NSE staining was positive.