Use of Perfluorodecalin for Pancreatic Islet Culture Prior to Transplantation: A Liquid-Liquid Interface Culture System—Preliminary Report

Abstract

Although the issue remains controversial, short-term culture is probably beneficial for islet graft quality. However, significant islet loss is invariably observed. This is related to reduced survival of large islets, which is compromised by hypoxia under standard culture conditions. We aimed to develop a method of culture, which would avoid exposure to relative hypoxia and hence maintain the quality of islets. Isolated rat islets cultured for 48 h in a liquid-liquid interface culture system (LICS) with a perfluorocarbon were compared to islets cultured under standard (C1) and suboptimal conditions (C2). Islets were tested for viability and response to a glucose challenge, and a marginal mass was transplanted into syngeneic diabetic recipients. The viability of islets after 24-h culture in LICS was higher than in C1 and C2 groups (89.0% vs. 77.5% and 64.6%, respectively) and decreased with time to reach 79.0%, 62.9%, and 53.4% after 72-h culture. The stimulation index in LICS-cultured islets was also significantly higher than in C1 and C2 groups (12.3 ± 0.4 vs. 5.8 ± 0.5 and 4.1 ± 0.2, respectively). Following transplantation of LICS-cultured islets 50% of recipients were rendered normoglycemic compared with 14.3% and 31.3% for C2 and fresh islets, respectively. Our liquid-liquid interface culture system using perfluorodecalin provides optimized culture conditions, which preserve both islet viability and their ability to engraft successfully after intraportal transplantation and could be used for islet transportation.

Perfluorocarbon Emulsions Prevent Hypoxia of Pancreatic β-Cells

“…Although we have not provided direct evidence of better diffusion of oxygen to islets' core, a the medium improves function of pseudoislets cultured for 3 days compared with pseudoislets cultured in stan-lack of increase in VEGF expression in islets cultured in nonhyperoxygenated PFOB emulsion with concomitant dard conditions. We also found that expression of HIF-1α and VEGF increased significantly in pseudoislets preservation of their viability and secretory function may suggest that perfluorocarbon emulsion facilitates diffusion during this period of culture in standard conditions, whereas it remained stable in pseudoislets cultured in of oxygen to the islets even at normal oxygen partial pressures, as demonstrated previously (17). However, we can-PFOB supplemented medium.…”

Reduction of oxygen inhibition effect for microalgal growth using fluoroalkylated methoxy polyethylene glycol-stabilized perfluorocarbon nano-oxygen carriers

“…PFCs have been widely used in medical applications to deliver oxygen to mammalian tissues and/or remove metabolically waste as so-called blood substitutes [27,28]. Additionally, using PFCs as oxygen carriers to enhance the efficiency of microbial, plant or animal cell growth has also been extensively reported in the past two decades [29][30][31]. Previously, we have demonstrated the feasibility of using PFC liquids in twophase PFC/medium forms to enhance the growth of microalgae in a closed and hyperoxygenated spinner flask [32].…”

Current Status of Immunomodulatory and Cellular Therapies in Preclinical and Clinical Islet Transplantation

“…Significant islet loss related to reduced survival of large islets compromised by hypoxia has been observed under standard culture conditions. In order to improve the islet graft quality prior to transplantation, rat islets have been cultured for 48 h in a liquid-liquid interface culture system (LICS) using perfluorodecalin, a method of culture that avoids exposure of islets to relative hypoxia [209]. Results indicated that this protocol optimised culture conditions, which preserved both islet viability and significantly increased their ability to engraft successfully after intraportal transplantation and could be used for islet transportation.…”