Vinod, Elizabeth

James, Jithu Varghese

Kachroo, Upasana

Sathishkumar, Solomon

Livingston, Abel

Ramasamy, Boopalan

2019-10-01

In vivo tracking of labelled cells can provide valuable information about cellular behavior in the microenvironment, migration and contribution of transplanted cells toward tissue regeneration. Articular cartilage derived chondroprogenitors (CPs) show promise as a candidate for cell-based therapy as they have been classified as mesenchymal stem cells with inherent chondrogenic potential. Iron oxide labelling is known to withstand harsh processing techniques known to be associated with staining of osteochondral specimens. The aim of our study was to investigate the feasibility of labelling CPs with micron-sized super paramagnetic iron oxide (M-SPIO) particles and to study the effects of this approach on the labelling efficiency, viability, maintenance of phenotype and potential for differentiation. Human CPs were isolated using fibronectin adhesion assay, passage 2 cells were labelled using three concentrations of M-SPIO (12.75 μg/ml, 25.5 μg/ml and 38.25 μg/ml). At sub confluence, cells were assessed for a) iron uptake by Prussian blue stain and colorimetry b) viability using 7-amino actinomycin D, c) MSC marker expression by flow cytometric analysis and d) trilineage differentiation potential. Iron uptake was higher with increase in M-SPIO concentration whereas CD73, CD90 marker expression significantly decreased and chondrogenic potential appreciably reduced with increase in M-SPIO concentration. In conclusion, 12.75 μg/ml M-SPIO can successfully label human articular cartilage derived chondroprogenitors with minimal effect on cellular viability, MSC marker expression and potential for differentiation.

Department of Physiology, Christian Medical College, Vellore, 632002, India; Centre for Stem Cell Research, Christian Medical College, Vellore, 632002, India. Electronic address: elsyclarence@cmcvellore.ac.in..

Department of Biochemistry, Christian Medical College, Vellore, 632002, India. Electronic address: jithujames04@gmail.com..

Department of Physiology, Christian Medical College, Vellore, 632002, India. Electronic address: upasana_k@hotmail.com..

Department of Physiology, Christian Medical College, Vellore, 632002, India. Electronic address: solomon@cmcvellore.ac.in..

Department of Orthopaedics, Christian Medical College, Vellore, 632004, India. Electronic address: abel_livingston@cmcvellore.ac.in..

Department of Orthopaedics, Royal Darwin Hospital, Tiwi, NT, 0810, Australia. Electronic address: jpboopy@gmail.com..

Acta Histochem . 2019 Oct;121(7):791-797. doi: 10.1016/j.acthis.2019.07.004. Epub 2019 Jul 18.

https://pubmed.ncbi.nlm.nih.gov/31326114/?otool=iaurydwlib

https://hdl.handle.net/10137/7802

Chondroprogenitors

Decalcification

Iron labelling

MSC markers

Viability

Comparison of incremental concentrations of micron-sized superparamagnetic iron oxide for labelling articular cartilage derived chondroprogenitors.

Journal Article

Publication