The Preliminary Experimental Research On Trans-differentiation Of Peripheral Blood CD14~+Cells Into Corneal Endothelial Cells By Monocytes Migration Model

Objective: Explore an animal model to increase the number of the monocytes.Toimitate the model of transendothelial migration of peripheral blood CD14~+cells invitro,and explore the possibility of trans-differentiation after they co-cultured withcornea endothelial cells(CECs).Methods:1. Explore an animal model to increase the percentage of the monocytes byimplanting foreign body in rabbits’ abdomen.2.Isolate the peripheral blood CD14~+cells by density gradient centrifugationmethod and detect the cell amount by flow cytometry.3.Simulate the model in vitro that peripheral blood CD14~+cells migratethrough human umbilical vein endothelial cells,inverted microscope is used to observethe morphology of cells after migration, then induce the cells towards adipogenicdirection, and evaluated the differentiation competence by specific staining.4. Cells which were collected from the model were co-cultured withCECs，the morphology changes were studied by inverted microscope, and the AQP-1positive rate were measured by flow cytometry.Results:1.After surgical intervention with foreign body implantation,the percentagedifferences of monocytes are statistically significant.(P <0.05).2.After cultured in inducing medium,these cells which obtained from modeof transendothelial migration of peripheral blood CD14~+cells were successfullyinduced into adipogenic.They were positive for oil red O staining.3.Co-culture the cells from the transendothelial migration model withCECs,these cells’morphology gradually changed from long to triangle,similar circularor irregular shape.With flow cytometry, the positive rate of AQP-1is0.6%.Conclusions:1.The monocyte percentage after the foreign body implantationintervention improved significantly compared with that before the implantation. 2.The cells obtained from the transendothelial migration model can beinduced to differentiate into adipocytes in vitro.3.After co-cultured with CECs, the cells’morphology changed andflow cytometric shows that the positive rate of AQP-1was0.6%.Preliminary view that the peripheral blood CD14~+cells possibly have some functionof the CECs after the model of transendothelial migration,which is a necessarygroundwork for our follow-up experimental study.