Microfluidic device for multimodal characterization of pancreatic islets.

A microfluidic device to perfuse pancreatic islets while simultaneously characterizing their functionality through fluorescence imaging of the mitochondrial membrane potential and intracellular calcium ([Ca(2+)](i)) in addition to enzyme linked immunosorbent assay (ELISA) quantification of secreted insulin was developed and characterized. This multimodal characterization of islet function will facilitate rapid assessment of tissue quality immediately following isolation from donor pancreas and allow more informed transplantation decisions to be made which may improve transplantation outcomes. The microfluidic perfusion chamber allows flow rates of up to 1 mL min(-1), without any noticeable perturbation or shear of islets. This multimodal quantification was done on both mouse and human islets. The ability of this simple microfluidic device to detect subtle variations in islet responses in different functional assays performed in short time-periods demonstrates that the microfluidic perfusion chamber device can be used as a new gold standard to perform comprehensive islet analysis and obtain a more meaningful predictive value for islet functionality prior to transplantation into recipients, which is currently difficult to predict using a single functional assay.

[1]  Michael G. Roper,et al.  Islet secretory defect in insulin receptor substrate 1 null mice is linked with reduced calcium signaling and expression of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA)-2b and -3. , 2004, Diabetes.

[2]  F. Sundler,et al.  Local Growth Factors Are Beneficial for the Autonomic Reinnervation of Transplanted Islets in Rats , 2003, Pancreas.

[3]  Wendell A Lim,et al.  A microfluidic system for dynamic yeast cell imaging. , 2008, BioTechniques.

[4]  E. Ryan,et al.  Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. , 2000, The New England journal of medicine.

[5]  R. Gomis,et al.  Pulsatile insulin release from islets isolated from three subjects with type 2 diabetes. , 2002, Diabetes.

[6]  S. Theander,et al.  Beta-cell secretory products activate alpha-cell ATP-dependent potassium channels to inhibit glucagon release. , 2005, Diabetes.

[7]  O. Korsgren,et al.  The ADP/ATP Ratio: A Novel Predictive Assay for Quality Assessment of Isolated Pancreatic Islets , 2006, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[8]  Matthew D. Davis,et al.  Retinopathy and nephropathy in patients with type 1 diabetes four years after a trial of intensive therapy. , 2000, The New England journal of medicine.

[9]  L. Brown,et al.  Acetoacetate and beta-hydroxybutyrate in combination with other metabolites release insulin from INS-1 cells and provide clues about pathways in insulin secretion. , 2008, American journal of physiology. Cell physiology.

[10]  P. Bergsten,et al.  Preserved Pulsatile Insulin Release from Prediabetic Mouse Islets1. , 1999, Endocrinology.

[11]  S. Sigrist,et al.  Induction of Angiogenesis in Omentum with Vascular Endothelial Growth Factor: Influence on the Viability of Encapsulated Rat Pancreatic Islets during Transplantation , 2003, Journal of Vascular Research.

[12]  J. Miyazaki,et al.  Hyperplastic islets observed in "reversed" NOD mice treated without hematopoietic cells. , 2008, Diabetes research and clinical practice.

[13]  R. Kennedy,et al.  On‐line competitive immunoassay based on capillary electrophoresis applied to monitoring insulin secretion from single islets of Langerhans , 1998, Electrophoresis.

[14]  David W Piston,et al.  Microfluidic glucose stimulation reveals limited coordination of intracellular Ca2+ activity oscillations in pancreatic islets. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Gilon,et al.  Oscillations of secretion driven by oscillations of cytoplasmic Ca2+ as evidences in single pancreatic islets. , 1993, The Journal of biological chemistry.

[16]  B. Soria,et al.  Glucose-induced [Ca2+]i oscillations in single human pancreatic islets. , 1996, Cell calcium.

[17]  C. Ricordi,et al.  Adenoviral gene transfer of erythropoietin confers cytoprotection to isolated pancreatic islets , 2004, Transplantation.

[18]  C. Ricordi,et al.  Twenty years of clinical islet transplantation at the Diabetes Research Institute--University of Miami. , 2004, Clinical transplants.

[19]  John F. Dishinger,et al.  Serial immunoassays in parallel on a microfluidic chip for monitoring hormone secretion from living cells. , 2007, Analytical chemistry.

[20]  Y. Bae,et al.  Visual evidence and quantification of interaction of polymeric sulfonylurea with pancreatic islet. , 2003, Biomacromolecules.

[21]  L. Philipson,et al.  Improved Outcomes in Islet Isolation and Transplantation by the Use of a Novel Hemoglobin‐based O2 Carrier , 2006, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[22]  S. Theander,et al.  β-Cell Secretory Products Activate α-Cell ATP-Dependent Potassium Channels to Inhibit Glucagon Release , 2005 .

[23]  R. Bretzel,et al.  Ischaemia is linked to inflammation and induction of angiogenesis in pancreatic islets , 2006, Clinical and experimental immunology.

[24]  D. Sutherland,et al.  Pancreas transplant outcomes for United States (US) and non-US cases as reported to the United Network for Organ Sharing (UNOS) and the International Pancreas Transplant Registry (IPTR) as of October 2002. , 2002, Clinical transplants.

[25]  B. Soria,et al.  The relationship between glucose‐induced K+ATP channel closure and the rise in [Ca2+]i in single mouse pancreatic beta‐cells. , 1992, The Journal of physiology.

[26]  Louis Scampavia,et al.  Micro sequential injection: automated insulin derivatization and separation using a lab-on-valve capillary electrophoresis system. , 2003, The Analyst.

[27]  P. Bergsten,et al.  Preserved pulsatile insulin release from prediabetic mouse islets. , 1999, Endocrinology.

[28]  C. Ricordi,et al.  The Use of Multiparametric Monitoring during Islet Cell Isolation and Culture: A Potential Tool for In-Process Corrections of Critical Physiological Factors , 2004, Cell transplantation.

[29]  Michael G. Roper,et al.  Microfluidic chip for continuous monitoring of hormone secretion from live cells using an electrophoresis-based immunoassay. , 2003, Analytical chemistry.

[30]  M. Mcdaniel,et al.  Hyperinsulinism in mice with heterozygous loss of KATP channels , 2006, Diabetologia.

[31]  G. Patterson,et al.  Separation of the glucose-stimulated cytoplasmic and mitochondrial NAD(P)H responses in pancreatic islet beta cells. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[32]  R T Kennedy,et al.  On-line competitive immunoassay for insulin based on capillary electrophoresis with laser-induced fluorescence detection. , 1996, Analytical chemistry.

[33]  J. Stockman.  Insulin Needs After CD3-Antibody Therapy in New-Onset Type 1 Diabetes , 2007 .

[34]  Wei-Jun Qian,et al.  Simultaneous monitoring of Zn2+ secretion and intracellular Ca2+ from islets and islet cells by fluorescence microscopy. , 2004, BioTechniques.

[35]  J. García-Sancho,et al.  Fluorescence Digital Image Analysis of Glucose-Induced [Ca2+]i Oscillations in Mouse Pancreatic Islets of Langerhans , 1993, Diabetes.

[36]  H. Vaudry,et al.  Pituitary Adenylate Cyclase-Activating Polypeptide , 2003, Endocrine Updates.

[37]  David W Piston,et al.  Critical Role of Gap Junction Coupled KATP Channel Activity for Regulated Insulin Secretion , 2006, PLoS biology.

[38]  Yong Wang,et al.  Human islet isolation outcomes from pancreata preserved with Histidine-Tryptophan Ketoglutarate versus University of Wisconsin solution. , 2006, Transplantation.

[39]  S. Carobbio,et al.  In beta-cells, mitochondria integrate and generate metabolic signals controlling insulin secretion. , 2006, The international journal of biochemistry & cell biology.

[40]  R. Martín del Río,et al.  Conversion into GABA (gamma-aminobutyric acid) may reduce the capacity of L-glutamine as an insulin secretagogue. , 2004, The Biochemical journal.

[41]  K. Polonsky,et al.  A 48-hour exposure of pancreatic islets to calpain inhibitors impairs mitochondrial fuel metabolism and the exocytosis of insulin. , 2003, Metabolism: clinical and experimental.

[42]  D. Sutherland,et al.  Pancreas transplant outcomes for United States (US) cases as reported to the United Network for Organ Sharing (UNOS) and the International Pancreas Transplant Registry (IPTR). , 2008, Clinical transplants.

[43]  W. Zawalich,et al.  Effects of protein kinase C inhibitors on insulin secretory responses from rodent pancreatic islets , 2001, Molecular and Cellular Endocrinology.

[44]  P. Casey,et al.  Gαz Negatively Regulates Insulin Secretion and Glucose Clearance* , 2008, Journal of Biological Chemistry.

[45]  P. Gilon,et al.  Distinct effects of glucose on the synchronous oscillations of insulin release and cytoplasmic Ca2+ concentration measured simultaneously in single mouse islets. , 1995, Endocrinology.

[46]  Robert T Kennedy,et al.  Perfusion and chemical monitoring of living cells on a microfluidic chip. , 2005, Lab on a chip.

[47]  W. Qian,et al.  Microscale determination of purines in tissue samples by capillary liquid chromatography with electrochemical detection. , 2003, The Analyst.

[48]  T. Ishikawa,et al.  Constitutive nitric oxide synthases in rat pancreatic islets: direct imaging of glucose-induced nitric oxide production in β-cells , 2003, Pflügers Archiv.

[49]  I. Todorov,et al.  Glucose‐Stimulated Increment in Oxygen Consumption Rate as a Standardized Test of Human Islet Quality , 2007, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[50]  Blue Cross.  Islet Transplantation in Patients with Type 1 Diabetes Mellitus , 2004 .

[51]  B. Soria,et al.  Glucoseminduced [Ca2+]i oscillations in single human pancreatic islets , 1996 .

[52]  A. Steffen,et al.  Quantification of Basal and Stimulated ROS Levels as Predictors of Islet Potency and Function , 2007, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[53]  G. Weir,et al.  A stirred microchamber for oxygen consumption rate measurements with pancreatic islets , 2007, Biotechnology and bioengineering.

[54]  J. Deeney,et al.  The L-type voltage-gated Ca2+ channel is the Ca2+ sensor protein of stimulus-secretion coupling in pancreatic beta cells. , 2007, Biochemistry.