Macedonian Veterinary Review

Original Scientific Article

In-vitro labelling of ovine adipose-derived mesenchymal stem cells (oADMSCS) and tracking using MRI technique

Ravi Gnanam Gnanadevi ,

Geetha Ramesh ,

Thandavan Arthanari Kannan * ,

Benjamin Justin William ,

Manoharan Parthiban ,

Gnanasigamani Sathyan

Mac Vet Rev 2017; 40 (2): 137 - 142

10.1515/macvetrev-2017-0018

Received: 27 November 2016

Received in revised form: 13 February 2017

Accepted: 15 April 2017

Available Online First: 22 May 2017

Published on: 15 October 2017

Correspondence: Thandavan Arthanari Kannan, kanns2000@gmail.com

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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.

Keywords: ovine, mesenchymal stem cells, in-vitro MIRB labelling, MRI imaging

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Copyright

© 2017 Gnanadevi R.G.This is an open-access article published under the terms of the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Conflict of Interest Statement

The authors declared that they have no potential conflict of interest with respect to the authorship and/or publication of this article.

Citation Information

Macedonian Veterinary Review. Volume 40, Issue 2, Pages 137-142, p-ISSN 1409-7621, e-ISSN 1857-7415, DOI: 10.1515/macvetrev-2017-0018, 2017