Study Of CD45~-/CD31~+ Mouse Lung Side Population Cells Develop Into Vascular Endothelial Cells And Smooth Muscle Cells In Vitro

ObsjectThe lung is an integral part of the respiratory system,its function is to allow the exchange of gas,the good lung function can maintain the protection of life.Pulmonary diseases are more common today and are very serious.Acute lung injury is a common type of disease in lung disease,which is caused by a variety of injuries,can cause pulmonary failure.Pulmonary disease seriously affects human health.In this study,CD45~-/CD31~+ mice lung side population cells were used to differentiate and induct in vitro,studying CD45~-/CD31~+mice lung side population cells may be the progenitor cells of the vascular endothelial cells and smooth muscle cells.Laying the foundation for treating the lung injury.Methods1.Collected the cells of lung tissue and sorted CD45~-/CD31~+ mice lung side population cells by flow cytometry.Immunofluorescence detected the expression of ABCG2.2.RT-PCR detected the expression of CD45~-/CD31~+ mice lung side population cells vascular endothelial cell related genes(Tie2,v WF)and smooth muscle cell related genes(SMA,?-SMT).3.The sorted cells were confirmed it is CD45~-/CD31~+ mice lung side population cells with cell clone formation experiments and flow cytometry.4.Cultured CD45~-/CD31~+ mice lung side population cells respectively with SmGm-2 and EGM-2,immunofluorescence detected the expressed of ?-SMT and v WF.5.The expression of smooth muscle cell-related genes ?-SMT and SMA,and the expression of vascular endothelial cell-associated gene v WF in CD45~-/CD31~+ mice lungs induced by in vitro differentiation and not induced were detected by RT-PCR.Results1.Flow cytometry successfully sorted CD45~-/CD31~+mice lung side population cells,and immunofluorescence detected taht it express ABCG2.2.Endothelial cell related gene test results showed that it expressed Tie2,but v WF was not expressed;Smooth muscle cell related gene test results showed it expressed SMA,but ?-SMT was not expressed.3.Cell clone formation experiment was successful,flow cytometry technology verify that it is CD45~-/CD31~+ mice lung side population cells.4.Immunofluorescence confirmed that SmGm-2 cultured cells expressed?-SMT and EGM-2 cultured cells expressed v WF.5.RT-PCR confirmed that SmGm-2 cultured cells expressed ?-SMT,SMA,did not express ABCG2;EGM-2 cultured cells expressed v WF,did not express ABCG2.ConclusionsCD45~-/CD31~+ mice lung side population cells can develop into vascular endothelial cells and smooth muscle cells in vitro,suggesting that CD45~-/CD31~+ mice lung side population cells may be the progenitor cells ofvascular endothelial cells and smooth muscle cell.