A stirred microchamber for oxygen consumption rate measurements with pancreatic islets.
Improvements in pancreatic islet transplantation for treatment of diabetes are hindered by the absence of meaningful islet quality assessment methods. Oxygen consumption rate (OCR) has previously been used to assess the quality of organs and primary tissue for transplantation. In this study, we describe and characterize a stirred microchamber for measuring OCR with small quantities of islets. The device has a titanium body with a chamber volume of about 200 microL and is magnetically stirred and water jacketed for temperature control. Oxygen partial pressure (pO(2)) is measured by fluorescence quenching with a fiber optic probe, and OCR is determined from the linear decrease of pO(2) with time. We demonstrate that measurements can be made rapidly and with high precision. Measurements with betaTC3 cells and islets show that OCR is directly proportional to the number of viable cells in mixtures of live and dead cells and correlate linearly with membrane integrity measurements made with cells that have been cultured for 24 h under various stressful conditions.
Papas
Klearchos K
KK
Department of Chemical Engineering, Massachusetts Institute of Technology, 25 Ames St., Cambridge, Massachusetts 02139, USA.
Pisania
Anna
A
Wu
Haiyan
H
Weir
Gordon C
GC
Colton
Clark K
CK
D000465:Algorithms; D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D016381:Islets of Langerhans Transplantation / Q000379:methods; D056656:Lab-On-A-Chip Devices; D008297:Male; D051379:Mice; D046208:Microchip Analytical Procedures / Q000379:methods; D010100:Oxygen / Q000032:analysis / Q000378:metabolism; D010101:Oxygen Consumption; D010313:Partial Pressure; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013552:Swine
D010100:Oxygen