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      Improved Labeling of Pancreatic Islets Using Cationic Magnetoliposomes

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          Abstract

          Pancreatic islets (PIs) transplantation is an alternative approach for the treatment of severe forms of type 1 diabetes (T1D). To monitor the success of transplantation, it is desirable to follow the location of engrafted PIs non-invasively. In vivo magnetic resonance imaging (MRI) of transplanted PIs is a feasible cell tracking method; however, this requires labeling with a suitable contrast agent prior to transplantation. We have tested the feasibility of cationic magnetoliposomes (MLs), compared to commercial contrast agents (Endorem and Resovist), by labeling insulinoma cells and freshly isolated rat PIs. It was possible to incorporate Magnetic Ressonance (MR)-detectable amounts of MLs in a shorter time (4 h) when compared to Endorem and Resovist. MLs did not show negative effects on the PIs’ viability and functional parameters in vitro. Labeled islets were transplanted in the renal sub-capsular region of healthy mice. Hypointense contrast in MR images due to the labeled PIs was detected in vivo upon transplantation, while MR detection of PIs labeled with Endorem and Resovist was only possible after the addition of transfection agents. These findings indicate that MLs are suitable to image PIs, without affecting their function, which is promising for future longitudinal pre-clinical and clinical studies involving the assessment of PI transplantation.

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          Glucose sensitivity and metabolism-secretion coupling studied during two-year continuous culture in INS-1E insulinoma cells.

          Rat insulinoma-derived INS-1 cells constitute a widely used beta-cell surrogate. However, due to their nonclonal nature, INS-1 cells are heterogeneous and are not stable over extended culture periods. We have isolated clonal INS-1E cells from parental INS-1 based on both their insulin content and their secretory responses to glucose. Here we describe the stable differentiated INS-1E beta-cell phenotype over 116 passages (no. 27-142) representing a 2.2-yr continuous follow-up. INS-1E cells can be safely cultured and used within passages 40-100 with average insulin contents of 2.30 +/- 0.11 microg/million cells. Glucose-induced insulin secretion was dose-related and similar to rat islet responses. Secretion saturated with a 6.2-fold increase at 15 mm glucose, showing a 50% effective concentration of 10.4 mm. Secretory responses to amino acids and sulfonylurea were similar to those of islets. Moreover, INS-1E cells retained the amplifying pathway, as judged by glucose-evoked augmentation of insulin release in a depolarized state. Regarding metabolic parameters, INS-1E cells exhibited glucose dose-dependent elevations of NAD(P)H, cytosolic Ca(2+), and mitochondrial Ca(2+) levels. In contrast, mitochondrial membrane potential, ATP levels, and cell membrane potential were all fully activated by 7.5 mm glucose. Using the perforated patch clamp technique, 7.5 and 15 mm glucose elicited electrical activity to a similar degree. A K(ATP) current was identified in whole cell voltage clamp using diazoxide and tolbutamide. As in native beta-cells, tolbutamide induced electrical activity, indicating that the K(ATP)conductance is important in setting the resting potential. Therefore, INS-1E cells represent a stable and valuable beta-cell model.
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            Optimal implantation site for pancreatic islet transplantation.

            Since the first report of successful pancreatic islet transplantation to reverse hyperglycaemia in diabetic rodents, there has been great interest in determining the optimal site for implantation. Although the portal vein remains the most frequently used site clinically, it is not ideal. About half of the islets introduced into the liver die during or shortly after transplantation. Although many patients achieve insulin independence after portal vein infusion of islets, in the long term most resume insulin injections. This review considers possible sites and techniques of islet transplantation in small and large animal models, and in humans. Metabolic, immunological and technical aspects are discussed. Many groups have sought an alternative site that might offer improved engraftment and long-term survival, together with reduced procedure-related complications. The spleen, pancreas, kidney capsule, peritoneum and omental pouch have been explored. The advantages and disadvantages of various sites are discussed in order to define the most suitable for clinical use and to direct future research.
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              Nanoparticles for cell labeling.

              Cell based therapeutics are emerging as powerful regimens. To better understand the migration and proliferation mechanisms of implanted cells, a means to track cells in living subjects is essential, and to achieve that, a number of cell labeling techniques have been developed. Nanoparticles, with their superior physical properties, have become the materials of choice in many investigations along this line. Owing to inherent magnetic, optical or acoustic attributes, these nanoparticles can be detected by corresponding imaging modalities in living subjects at a high spatial and temporal resolution. These features allow implanted cells to be separated from host cells; and have advantages over traditional histological methods, as they permit non-invasive, real-time tracking in vivo. This review attempts to give a summary of progress in using nanotechnology to monitor cell trafficking. We will focus on direct cell labeling techniques, in which cells ingest nanoparticles that bear traceable signals, such as iron oxide or quantum dots. Ferritin and MagA reporter genes that can package endogenous iron or iron supplement into iron oxide nanoparticles will also be discussed.
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                Author and article information

                Journal
                J Pers Med
                J Pers Med
                jpm
                Journal of Personalized Medicine
                MDPI
                2075-4426
                12 March 2018
                March 2018
                : 8
                : 1
                : 12
                Affiliations
                [1 ]Biomedical MRI/MoSAIC, Department of Imaging and Pathology, University of Leuven, Herestraat 49, B3000 Leuven, Belgium; Rita.Sofia.Garcia.Ribeiro@ 123456vub.ac.be (R.S.G.R.); ashket2982@ 123456gmail.com (A.K.-A.); yinting527@ 123456gmail.com (T.Y.)
                [2 ]Laboratory for Functional and Metabolic Imaging, Ecole Polytechnique Federale Lausanne, 1015 Lausanne, Switzerland
                [3 ]Laboratory of Experimental Hormonology, ULB, Campus Erasme, Route de Lennik 808, B1070 Brussels, Belgium; karim.louchami@ 123456ulb.ac.be (K.L.); abdsener@ 123456ulb.ac.be (A.S.)
                [4 ]Laboratory of Histology, Biomedical Research Institute, Hasselt University, Campus Diepenbeek, Agoralaan, B3590 Diepenbeek, Belgium; tom.struys@ 123456sprofit.com (T.S.); ivo.lambrichts@ 123456uhasselt.be (I.L.)
                [5 ]Department of Biochemistry, ULB, Campus Erasme, Route de Lennik 808, B1070 Brussels, Belgium; malaisse@ 123456ulb.ac.be
                [6 ]Laboratory of BioNanoColloids, Interdisciplinary Research Centre, University of Leuven, Etienne Sabbelaan 53, B8500 Kortrijk, Belgium; marcel.decuyper@ 123456kuleuven.be
                Author notes
                [* ]Correspondence: Uwe.Himmelreich@ 123456med.kuleuven.be ; Tel.: +32-16-330-925
                [†]

                These authors contributed equally to this manuscript.

                Author information
                https://orcid.org/0000-0001-7520-0021
                Article
                jpm-08-00012
                10.3390/jpm8010012
                5872086
                29534555
                ae8c6f68-bac9-4d9d-a5fe-c9299b2dd883
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 21 December 2017
                : 08 March 2018
                Categories
                Article

                pancreatic islets,islet transplantation,mri,contrast agents,magnetoliposomes,spios,insulin,diabetes mellitus,cell imaging,transmission electron microscopy

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