In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue |
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| Institution: | 1. Department of Transplant Biology, Immunology & Stem Cell Lab, Global Hospitals, Hyderabad, India;2. KK Nuclear Scans, Raj Bhavan Road, Somajiguda, Hyderabad, India;3. Cancer Centre, GA Regents University, Augusta, GA, USA;1. Department of Surgical Oncology, Cancer Treatment Center, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China;2. Department of Pharmacology and Toxicology, School of Pharmacy, Rutgers University, Piscataway, NJ, USA;3. Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, China;4. Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS, USA;5. Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS, USA;6. Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, MI, USA;7. Department of Medicine, Huntsman Cancer Institute, University of Utah School of Medicine, Salt Lake City, UT, USA;1. Mass spectrometry/Proteomics research Group, Max Planck Institute for Chemical Ecology, Hans-Knöll-Straße 8, D-07745 Jena, Germany;2. Department of Cell and Molecular Biology, Leibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Beutenbergstr. 11a, D-07745 Jena, Germany;3. Friedrich-Schiller-University Jena, D-07737 Jena, Germany;1. PET Center, Aarhus University Hospital, DK-8000 Aarhus C, Denmark;2. DanPET AB, Rosenstigen 7, SE-216 19 Malmö, Sweden;3. NeuroSearch A/S, Pederstrupsvej 93, DK-2750 Ballerup, Denmark |
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| Abstract: | IntroductionNoninvasive radionuclide imaging of cells using technetium99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) is a potential diagnostic tool for several applications. Herein we aimed to evaluate the labeling efficiency and cellular toxicity of 99mTc-HMPAO with Stromal Vascular Fraction (SVF) of adipose tissue to develop a process tool for theranostic purposes, in particular imaging cardiac stem cell therapy.MethodsTen million cells of SVF were labeled with 99mTc-HMPAO complex and excess radiolabel was cleared off through washing in PBS. The labeling efficiency of 99mTc-HMPAO was detected in labeled cells and their subsequent supernatant wash using isotope dose calibrator and gamma camera. The cytotoxicity was assessed for the comparative reactive oxygen species (ROS) by H2DCFDDA, apoptotic events by annexin-V and TUNEL assay and mitochondrial potential by JC-1.ResultsAn encouraging labeling efficiency of 33% was observed with 99mTc-HMPAO complex. The radionuclide labeling of SVF demonstrated significant safety profile as evaluated by apoptotic assays.Conclusion99mTc-HMPAO labeling efficiency of 33% of total SV fraction would produce sufficient radioactive signals that would enable for in vivo tracking of cells by SPECT-CT. The radionuclide did not demonstrate any significant impact on the structural or functional organization of the labeled cells. Our study indicates that SVF can be safely labeled with 99mTc-HMPAO without adverse cytotoxic events and for its potential role in imaging cardiac stem cell therapy. |
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