In-vitro Labelling of Ovine Adipose-Derived Mesenchymal Stem Cells (oADMSCs) and Tracking Using MRI Technique

Abstract

To understand the mechanisms standing behind a successful stem cell-based therapy, the monitoring of transplanted cell’s migration, homing as well as the engraftment efficiency and functional capability in-vivo has become a critical issue. The present study was designed to track the labelled oADMSCs in-vitro and its visualization through MRI technique. oADMSCs from passage 4 (P-4) to passage 6 (P-6) were labelled with superparamagnetic iron oxide (SPIO) conjugated with rhodamine (Molday Ion Rhodamine-B - MIRB) at the concentration of 25μg Fe/ml in DMEM. Internalized MIRB was observed under fluorescent microscope after 72 hrs of incubation. Labelled oADMSCs showed Prussian Blue positive reaction demonstrating the iron uptake of the cells. The viability of the MIRB-labelled oADMSCs ranged between 98-99 per cent and Trypan blue exclusion test showed no significant difference in viability between labelled and unlabelled oADMSCs. MR signal in control group of cells was similar to that of water. MR signals or fluorescence in MIRB-labelled cells decreased with increasing concentrations of iron. The T2 weighted images of MIRB-labelled oADMSCs increased with increasing concentrations of SPIOs. The MIRB was found to be nontoxic, and did not affect proliferation capacity in-vitro.