Study On The Surface Labeling Of The Bone Marrow Mesenchymal Stem Cell By Biotin-streptavidin

Objective1, Through the biotin streptavidin reaction system to establish a simple and feasible method for the modifying of cell surface.2, Evaluate the labeling rate of chemical modification method for the modified of mesenchymal stem cell.3, detect the effect of this chemical covalent binding on the biological function of bone marrow mesenchymal stem cells, founded the testing basis for this technology used in the promotion of stem cell homing in AKI.Method:1, The culture of whole bone marrow cells cultured bone marrow mesenchymal stem cells to third-generation spare,identificate of BMSCs by flow cytometry;2, biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were used to equip the adhesion molecule ligand, sialyated Lewis X(SLe X) to the BMSCs surface by the covalent blinding.3, The labeling rate of chemical modification method were identified by fluorescence microscope4, The effect of chemical modification method on the vitality of BMSCs were detected by Trypan blue staining5, The effect of chemical modification method on the proliferation of BMSCs were detected by CCK-8 assay6, Adipogenic, osteogenic induction were used to detect the effect of chemical modification methods on the multi differentiation function of BMSCs.Results:1,Whole bone marrow cells Can be extracted bone marrow mesenchymal stem cells, after culture 2 weeks, the third generation of BMSCs can be obtained,after identify by flow cytometry,the Cell surface marker conform to the phenotype of BMSC2,Evaluate the efficiency of chemical modification method: the modification efficiency of BNHS+S, was(87.8 + 3.1%), significantly higher than control group B+S(33.2 + 1.3)%(P<0.001) and group P+S(6.4 + 1.3%)(P<0.001);3,Evaluate the effect of chemical modification method on the biological characteristics of BMSCs: The 24 h and 48 h vitality of BMSCs after the processing BNHS were 83% and 76%, the result of CCK-8 assay show that the BNHS have little effect on the proliferation of BMSCs.23 days after addition of osteogenic differentiation media, PBS group and the BNHS modified group showed positive and similar staining for oil red O. 23 days after addition of adipogenic differentiation media, PBS group and the BNHS modified group showed showed positive and similar staining for Alizarin red O.Conclusion1, Through the biotin streptavidin reaction system, we equip the adhesion molecule ligand, sialyated Lewis X(SLe X) to the BMSCs surface by the covalent blinding,This approach is simple, has a mild reaction condition.2, The approach does not affect the differentiation of BMSC and has a high modification efficiency3, This method has great applicable valuest, may help to promote the homing of stem cells in acute kidney injury.