High-Density Culture of Human Islets on Top of Silicone Rubber Membranes

• Published in: Transplantation proceedings Vol. 37; no. 8; pp. 3412 - 3414
• Main Authors: Papas, K.K., Avgoustiniatos, E.S., Tempelman, L.A., Weir, G.C., Colton, C.K., Pisania, A., Rappel, M.J., Friberg, A.S., Bauer, A.C., Hering, B.J.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: New York, NY Elsevier Inc 01.10.2005; Elsevier Science
• Subjects: Biological and medical sciences; Cell Culture Techniques - methods; Cell Hypoxia; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; Islets of Langerhans - cytology; Islets of Langerhans - physiology; Medical sciences; Membranes, Artificial; Rubber; Silicones; Surface Properties; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Tissue, organ and graft immunology; High; Human; Langerhans islet; Transplantation; Density; Medicine; Treatment; Surgery; Graft; Membrane; Rubber; Siloxane polymer; Culture; Endocrine pancreas
• ISSN: 0041-1345; 1873-2623
• DOI: 10.1016/j.transproceed.2005.09.086

Islet culture has emerged as a standard practice prior to clinical transplantation. However, culturing large numbers of islets requires low islet density (number of islets per unit surface area) and, consequently, 20 to 30 flasks per pancreas in order to avoid hypoxia-induced death (HID). There is a need for a simple, practical, small-footprint culture vessel that will accommodate aseptic maintenance of entire human islet isolations while avoiding HID. In this communication, we examine the hypothesis that by improving oxygen transfer through culture of islets on silicone rubber membranes (SRM), we may increase islet surface coverage and reduce the number of flasks required while avoiding HID. Our results demonstrate that islets cultured for up to 48 hours in vessels with SRM bottoms at 2000 to 4000 islet equivalents (IE)/cm 2, a surface coverage 10- to 20-fold higher than the standard culture protocol, displayed no significant loss of viability. In contrast, islets cultured for 48 hours at 4000 IE/cm 2 in flasks with gas-impermeable bottoms suffered a 60% to 70% reduction in viability. The data suggest that it is possible to culture all islets isolated from a human pancreas on SRM in a single, standard-sized vessel while maintaining the same viability as with the current, standard culture protocols that require 20 to 30 flasks. This approach may lead to substantial improvements in islet culture for research and clinical transplantation.