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Homing and Tracking of Iron Oxide Labelled Mesenchymal Stem Cells After Infusion in Traumatic Brain Injury Mice: a Longitudinal In Vivo MRI Study

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Abstract

Stem cells transplantation has emerged as a promising alternative therapeutic due to its potency at injury site. The need to monitor and non-invasively track the infused stem cells is a significant challenge in the development of regenerative medicine. Thus, in vivo tracking to monitor infused stem cells is especially vital. In this manuscript, we have described an effective in vitro labelling method of MSCs, a serial in vivo tracking of implanted stem cells at traumatic brain injury (TBI) site through 7 T magnetic resonance imaging (MRI). Proper homing of infused MSCs was carried out at different time points using histological analysis and Prussian blue staining. Longitudinal in vivo tracking of infused MSCs were performed up to 21 days in different groups through MRI using relaxometry technique. Results demonstrated that MSCs incubated with iron oxide-poly-L-lysine complex (IO-PLL) at a ratio of 50:1.5 μg/ml and a time period of 6 h was optimised to increase labelling efficiency. T2*-weighted images and relaxation study demonstrated a significant signal loss and effective decrease in transverse relaxation time on day-3 at injury site after systemic transplantation, revealed maximum number of stem cells homing to the lesion area. MRI results further correlate with histological and Prussian blue staining in different time periods. Decrease in negative signal and increase in relaxation times were observed after day-14, may indicate damage tissue replacement with healthy tissue. MSCs tracking with synthesized negative contrast agent represent a great advantage during both in vitro and in vivo analysis. The proposed absolute bias correction based relaxometry analysis could be extrapolated for stem cell tracking and therapies in various neurodegenerative diseases.

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Fig. 1: Phenotypic and differential characterisation study of isolated mMSCs
Fig. 2
Fig. 3: Longitudinal in vivo tracking of unlabeled MSCs after administration in TBI mouse
Fig. 4: Longitudinal in vivo tracking of labeled MSCs after administration in TBI mouse
Fig. 5: Comparison of T2* time of injured area in between groups
Fig. 6: Fluorescent microscopy of injured area after infusion of labelled MSCs
Fig. 7: Prussian blue staining of injured area after infusion of labelled MSCs
Fig. 8: Histological analysis of injured area and its contralateral side after transplantation of dual labelled MSCs

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Abbreviations

ANOVA:

Analysis of variance

FBS:

Foetal Bovine Serum

FOV:

Field of view

ICC:

Immunocytochemistry

ISA:

Imaging sequence analysis

MGE:

Multi Gradient Echo

MRI:

Magnetic resonance imaging

MSCs:

Mesenchymal Stem Cells

mMSCs:

mice MSCs

MSME:

Multi Slice Multi Echo

PCR:

Polymerase chain reaction

PLL:

Poly-L-lysine

ROI:

Region of interest

R2 :

Transverse relaxation rate

T2 :

Transverse relaxation time

TR:

Repetition time

TE:

Echo time

TBI:

Traumatic brain injury

USPIO:

Ultrasmall superparamagnetic iron oxide

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Acknowledgements

The S & T project (INM-311, 4.1/1.6) was supported by funding from the Defence Research and Development Organization (DRDO), Ministry of Defence, Govt. of India. The authors also wish to express their gratitude to Dr. Anurag Agrawal and Bijay Ranjan Pattnaik (Research Fellow), Translational research in asthma laboratory, IGIB for their unstinted support in the microscopy study. Dr. Sushanta Kumar Mishra sincerely thanks the Indian Council of Medical Research (ICMR, 2014-25070) for providing a research fellowship in support of the project.

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Authors and Affiliations

  1. NMR Research Centre, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, -54, Delhi, India

    Sushanta Kumar Mishra & Subash Khushu

  2. Division of Stem Cell and Gene Therapy Research, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, -54, Delhi, India

    Sushanta Kumar Mishra, Ajay K. Singh & Gurudutta Gangenahalli

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  1. Sushanta Kumar Mishra
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  2. Subash Khushu
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Contributions

SKM conceived the study, designed experimental plan, performed experiments, collected and interpreted data and drafted the manuscript. SK and GG supervised all the experimental work, assisted in data interpretation, crosschecked the results and revised the manuscript. AKS took administrative part in providing research funding and permitted to perform experimental work in other labs. All authors read and approved the final draft of manuscript.

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Correspondence to Subash Khushu or Gurudutta Gangenahalli.

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Mishra, S.K., Khushu, S., Singh, A.K. et al. Homing and Tracking of Iron Oxide Labelled Mesenchymal Stem Cells After Infusion in Traumatic Brain Injury Mice: a Longitudinal In Vivo MRI Study. Stem Cell Rev and Rep 14, 888–900 (2018). https://doi.org/10.1007/s12015-018-9828-7

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