title: The use of the BD oxygen biosensor system to assess isolated human islets of langerhans: oxygen consumption as a potential measure of islet potency.
abstract: The measurement of cellular oxygen consumption rate (OCR) is a potential tool for the assessment of metabolic potency of isolated islets of Langerhans prior to clinical transplantation. We used a commercially available 96-well plate fluoroprobe, the BD Oxygen Biosensor System (OBS), to estimate OCR in 27 human islet preparations, and compared these results to those of concurrent mouse transplantations. OCR was estimated both from the dO2 at steady state and from the transient rate of change of dO2 during the initial culture period immediately after seeding ("dO2 slope"). To demonstrate the validity of the OBS-derived values, it was shown that they scaled linearly with islet equivalent number/DNA concentration and with each other. These measurements were obtained for each preparation of islets incubated in media supplemented with either low (2.2 mM) or high (22 mM) glucose. Concurrently, one to three athymic nude mice were transplanted with 2,000 IEQs under the kidney capsule. The OCR Index, defined as the ratio of the DNA-normalized "dO2 slope" in high glucose to that in low glucose, proved highly predictive of mouse transplant results. Of the 69 mice transplanted, those receiving islets where the OCR Index exceeded 1.27 were 90% likely to reverse within 3 days, whereas those receiving islets with an OCR Index below 1.27 took significantly longer, often failing to reverse at all over a 35-day time period. These results suggest that the OBS could be a useful tool for the pretransplant assessment of islet cell potency.
authors.affiliation: Diabetes Research Institute, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Fraker
authors.forename: Mark R
authors.initials: MR
authors.lastname: Timmins
authors.forename: Richard D
authors.initials: RD
authors.lastname: Guarino
authors.forename: Perry D
authors.initials: PD
authors.lastname: Haaland
authors.forename: Hirohito
authors.initials: H
authors.lastname: Ichii
authors.forename: Damaris
authors.initials: D
authors.lastname: Molano
authors.forename: Antonello
authors.initials: A
authors.lastname: Pileggi
authors.forename: Raffaella
authors.initials: R
authors.lastname: Poggioli
authors.forename: Sharon C
authors.initials: SC
authors.lastname: Presnell
authors.forename: Luca
authors.initials: L
authors.lastname: Inverardi
authors.forename: Mitra
authors.initials: M
authors.lastname: Zehtab
authors.forename: Camillo
authors.initials: C
authors.lastname: Ricordi
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D006085:Graft Survival / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D007515:Islets of Langerhans / Q000378:metabolism* / Q000502:physiology; D016381:Islets of Langerhans Transplantation / Q000379:methods; D016015:Logistic Models; D008297:Male; D051379:Mice; D008819:Mice, Nude; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption / Q000502:physiology; D014183:Transplantation, Heterologous
chemical_list: D010100:Oxygen