Title: Novel cell culture technology significantly improves islet function and practical advantage of good transportation of islets
Abstract: Event Abstract Back to Event Novel cell culture technology significantly improves islet function and practical advantage of good transportation of islets Sandeep Kumar1, 2, Nouf Alhasawi1, Claire Marriott1, Adrian Bone1 and Wendy Macfarlane1 1 University of Brighton, Pharmacy and Biomolecular Sciences, United Kingdom 2 Cellon SA, Luxembourg Aims: Pancreatic islet transplantation has emerged as a potential cure for type 1 diabetes. Human pancreatic islets are routinely cultured in static plastic tissue culture flask to ship from the isolation to the transplantation facility, which results into huge loss of islet mass and in vivo islet functionality. Our aim was to study the effect of enzymatic digestion on pseud islets extracellular matrix (ECM) and to determine if a microgravity rotary cell culture system (RCCS) will restore and enhance the ECM expression, viability and functionality of islets. Methods: Min6 beta cells were cultured as monolayers and then reconfigured into pseudoislet clusters (PIs) in either static dishes or a microgravity bioreactor. Specific functional markers, PDX1 and ECM proteins, were analysed by immunocytochemistry analysis (ICC). Specific ECM and insulin gene expression was analysed by qRT-PCR. PI digestion with collagenase was performed for a range of time points and cell viability was assessed (HPI assay). PIs were subsequently allowed to recover under static or bioreactor cell culture conditions. Results: PI cells cultured in both static and bioreactor conditions showed nuclear translocation of PDX1 after 1h of glucose stimulation. ECM gene and protein expression was altered in PIs maintained in static and bioreactor cultures. Fibronectin and laminin V expression was significantly increased under bioreactor compared with static conditions. Collagen IV was highly expressed in monolayers compared with PIs cultured in static and bioreactor. Insulin gene expression was markedly increased in bioreactor compared with static culture. Microscopic analysis showed an improved ECM restoration in PIs cultured in the bioreactor compared with static cultures. Conclusion: Our study has shown that microgravity cell culture has the potential to restore levels of ECM in islets following collagenase. The authors would like to thank Seventh Framework Programme. The authors would also like to thank University of Brighton, UK and Cellon SA, Luxembourg for funding and conducting this research work. Keywords: Extracellular Matrix, Gene Expression, Cell response, Cell functionality Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Modeling cellular events in tissue regeneration Citation: Kumar S, Alhasawi N, Marriott C, Bone A and Macfarlane W (2016). Novel cell culture technology significantly improves islet function and practical advantage of good transportation of islets. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.00158 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Sandeep Kumar Nouf Alhasawi Claire Marriott Adrian Bone Wendy Macfarlane Google Sandeep Kumar Nouf Alhasawi Claire Marriott Adrian Bone Wendy Macfarlane Google Scholar Sandeep Kumar Nouf Alhasawi Claire Marriott Adrian Bone Wendy Macfarlane PubMed Sandeep Kumar Nouf Alhasawi Claire Marriott Adrian Bone Wendy Macfarlane Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.