Use of perfluorodecalin for pancreatic islet culture prior to transplantation: a liquid-liquid interface culture system--preliminary report.
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.
Juszczak
M T
MT
Department of Endocrinology and Diabetes, Royal Free Hospital, Hempsted, London, UK. m.juszczak@doctors.net.uk
Elsadig
A
A
Kumar
A
A
Muzyamba
M
M
Pawelec
K
K
Powis
S H
SH
Press
M
M
D000165:Acridine Orange / Q000378:metabolism; D000818:Animals; D001681:Biological Assay; D001786:Blood Glucose / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D005215:Fasting / Q000097:blood; D005453:Fluorescence; D005466:Fluorocarbons / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D016381:Islets of Langerhans Transplantation; D009924:Organ Culture Techniques / Q000379:methods*; D010100:Oxygen; D010313:Partial Pressure; D011419:Propidium / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014022:Tissue Survival / Q000187:drug effects
D001786:Blood Glucose; D003470:Culture Media; D005466:Fluorocarbons; D011419:Propidium; C008805:perfluorodecalin; D000165:Acridine Orange; D010100:Oxygen