Superparamagnetic Iron Oxide And DAPI Double-labeling Bone Marrow Mesenchymal Stem Cells

Objective Now,bone marrow mesenchymal stem cells(BMSCs) has become important seed cells of organizational project. It is necessary to solve the problem that how to labled MSCs efficiently and safely before lucubrate MSCs' proliferation and division in vitro and in vivo.To investigate the effects of SPIO and DAPI double-labeling on bone marrow mesenchymal stem cells, and research whether there is any effect on cellular viability,proliferation and apoptosis and to determine the best marking method and lay the foundation for stem cells in vivo tracing study.Methods BMSCs were isolated and double-labeled with SPIO and DAPI, The labeling rate were identified by Prussian blue staining and fluorescence microscope, then detected it's vitality by Trypan blue staining and proliferation activity by MTT assay, the cell viability and apoptosis was observed by using Calceein AM/PI and AO/PI staining.Results The stem cell double-labeling rate by SPIO and DAPI were nearly 100%. The cell vitality rate with Trypan blue staining was 97%. MTT method detected that there was no significant difference for proliferation activity between double-labeled stem cells and those not labeled(P>0.05). Calcein-AM/PI staining detected that the survival rate of double-labeled and unlabeled stem cells were 95% and 96%. AO/PI staining detected that the apoptosis rate of double-labeled and unlabeled stem cells were both 1%.Conclusion There is no effect on survival,proliferation and apoptosis of bone marrow mesenchymal stem cells after it is labeled by superparamagnetic iron oxide and 4',6-diamidino-2-phenylindole.