Automated, high-throughput assays for evaluation of human pancreatic islet function.
An important challenge in pancreatic islet transplantation in association with type 1 diabetes is to define automatic high-throughput assays for evaluation of human islet function. The physiological techniques presently used are amenable to small-scale experimental samples and produce descriptive results. The postgenomic era provides an opportunity to analyze biological processes on a larger scale, but the transition to high-throughput technologies is still a challenge. As a first step to implement high-throughput assays for the study of human islet function, we have developed two methodologies: multiple automated perifusion to determine islet hormone secretion and high-throughput kinetic imaging to examine islet cellular responses. Both technologies use fully automated devices that allow performing simultaneous experiments on multiple islet preparations. Our results illustrate that these technologies can be applied to study the functional status and explore the pharmacological profiles of islet cells. These methodologies will enable functional characterization of human islet preparations before transplantation and thereby provide the basis for the establishment of predictive tests for beta-cell potency.
Cabrera
Over
O
Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, FL 33136, USA.
Jacques-Silva
M Caroline
MC
Berman
Dora M
DM
Fachado
Alberto
A
Echeverri
Felipe
F
Poo
Ramon
R
Khan
Aisha
A
Kenyon
Norma S
NS
Ricordi
Camillo
C
Berggren
Per-Olof
PO
Caicedo
Alejandro
A
D000818:Animals; D001322:Autoanalysis; D002118:Calcium / Q000032:analysis; D003924:Diabetes Mellitus, Type 2 / Q000503:physiopathology; D005934:Glucagon / Q000378:metabolism; D005947:Glucose / Q000032:analysis; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism / Q000502:physiology*; D016381:Islets of Langerhans Transplantation; D019520:Living Donors; D008252:Macaca fascicularis; D010187:Pancreatic Hormones / Q000378:metabolism; D010477:Perfusion; D013045:Species Specificity
D007328:Insulin; D010187:Pancreatic Hormones; D005934:Glucagon; D005947:Glucose; D002118:Calcium