Abstract:

BACKGROUND: Now, bone marrow mesenchymal stem cells (BMSCs) have become important seed cells of tissue engineering. It is necessary to solve the problem that how to label BMSCs efficiently and safely for further study on BMSCs proliferation and differentiation in vitro and in vivo.
OBJECTIVE: To investigate the effects of superparamagnetic iron oxide (SPIO) and 4',6-diamidino-2-phenylindole (DAPI) double-labeling on BMSCs, and to study whether there is any effect on cell viability, proliferation and apoptosis.
METHODS: BMSCs were isolated and double-labeled with SPIO and DAPI. The labeling rate was identified by Prussian blue staining and fluorescence microscope, then cell vitality was detected by Trypan blue staining and proliferation activity was measured by MTT assay. Cell survival rate and apoptosis rate were observed by using Calceein AM/PI and AO/PI staining.
RESULTS AND CONCLUSION: The stem cell double-labeling rate by SPIO and DAPI were nearly 100%. The cell survival rate with Trypan blue staining was 97%. MTT method detected that there was no significant difference for proliferation activity between double-labeled and unlabeled stem cells (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%. These suggested that there is no effect on survival, proliferation and apoptosis of BMSCs after SPIO and DAPI double-labeling.