Metabolic Tissue Function Core
A major goal of the core is to assist DRC investigators in obtaining metabolic tissues, including β-cell lines, rodent and human primary islets, and iPSCs for differentiation into diabetes relevant tissues as well as β-cells. The Core will also assist investigators in conducting functional and metabolic analysis of cells and tissues relevant to the pathogenesis of diabetes. These goals are accomplished through consultations, provision of expert service, and access to state-of-the-art equipment.
Diabetes mellitus comprises a group of disorders characterized by an absolute or relative deficiency of insulin. Diabetes can be associated with impaired function in many tissues regulating metabolism, including pancreatic β-cells, adipose tissue, skeletal muscle, and the hypothalamus. Many issues fundamental to understanding the etiology and consequences of diabetes are best understood at the level of tissue pathology, cellular structure and function. The purpose of the Metabolic Tissue Function Core is to improve the efficiency of diabetes-related research by providing services related to the structural and functional analyses of tissues important in the pathogenesis and complications of diabetes. A major goal of the core is to assist DRC investigators in obtaining metabolic tissues, including β-cell lines, rodent and human primary islets, and iPSCs for differentiation into diabetes relevant tissues as well as β-cells. The Core will also assist investigators in conducting functional and metabolic analysis of cells and tissues relevant to the pathogenesis of diabetes. These goals are accomplished through consultations, provision of expert service, and access to state-of-the-art equipment.
Leadership
Core Director
Fumihiko Urano, MD, PhD
Samuel E. Schechter Professor of Medicine, Professor of Medicine and Pathology & Immunology, Division of Endocrinology, Metabolism and Lipid Research
- Phone: 314- 362-8683
- Fax: 314-362-8770
- Email: urano@nospam.wustl.edu
Associate Director
Maria S. Remedi, PhD
Professor of Medicine and Cell Biology & Physiology, Division of Endocrinology, Metabolism, and Lipid Research
- Phone: 314-362-6636
- Fax: 314-362-7641
- Email: mremedi@nospam.wustl.edu
Core Manager
Cris Brown
Core Manager | Division of Endocrinology, Metabolism & Lipid Research
- Phone: 314-362-8684
- Email: crisbrown@nospam.wustl.edu
Research Lab Manager
Sarah Gale
Research Lab Manager | Division of Endocrinology, Metabolism & Lipid Research
- Email: segale@nospam.wustl.edu
Services
Metabolic Tissue Function Core
Diabetes mellitus comprises a group of disorders characterized by an absolute or relative deficiency of insulin. Diabetes can be associated with impaired function in many tissues regulating metabolism, including pancreatic β-cells, adipose tissue, skeletal muscle, and the hypothalamus. Many issues fundamental to understanding the etiology and consequences of diabetes are best understood at the level of tissue pathology, cellular structure and function. The purpose of the Metabolic Tissue Function Core is to improve the efficiency of diabetes-related research by providing services related to the structural and functional analyses of tissues important in the pathogenesis and complications of diabetes. A major goal of the core is to assist DRC investigators in obtaining metabolic tissues, including β-cell lines, rodent and human primary islets, and iPSCs for differentiation into diabetes relevant tissues as well as β-cells. The Core will also assist investigators in conducting functional and metabolic analysis of cells and tissues relevant to the pathogenesis of diabetes. These goals are accomplished through consultations, provision of expert service, and access to state-of-the-art equipment.
Metabolic Tissue Acquisition
- Advice for design of studies using islets or β-cells: Drs. Urano and Remedi provide consultations to researchers who are interested in using islets or β-cells but lack experience working with or generating these metabolic tissues.
- Rodent islet isolation: The Core assists DRC investigators in the isolation of primary islets from mouse models of diabetes and metabolic disorders. The procedure can be performed by Core personnel or they can provide training to personnel from DRC member labs, so that they can carry out their own isolations.
- Procurement of human primary islets: The Core assists investigators in preparing effective applications to the NIDDK-sponsored Integrated Islet Distribution Program (IIDP).
- Supply of β-cell lines: The Core maintains stocks of established β-cell lines, including MIN6, RINm5F, INS-1, βTC and βTC6. Upon request, investigators receive a frozen vial and standard operating procedures for culture.
- Generation of iPSC and genetically-modified cell lines: The Core leverages the well-established, high functioning Washington University Genome Engineering and iPSC Center (GEiC) to obtain iPSCs and genetically-modified cell lines for study of diabetes and its complications. DRC members are eligible for a one-time $1000 scholarship toward a diabetes-related project.
Metabolic Tissue Analyses
- Static Islet Secretory Responses: For rapid and cost-effective screening of insulin, glucagon, and somatostatin secretion in response to secretagogues, the Core offers static incubations in a 24 well plate format. Hormones are typically quantified by ELISA.
- Dynamic perifusion testing for quantification of physiological function of islets: Dynamic perifusion testing provides the most accurate assessment of regulated hormonal (i.e. insulin and glucagon) release ex-vivo to monitor islet function and physiology and enables pharmacological manipulation to dissect out signaling mechanisms and kinetics (e.g., first and second phase insulin secretion, pulsatile glucagon secretion) underlying release of different hormones.
- Islet Morphometry: The Core provides rigorous quantification of β-cell mass using histological staining and systematic imaging and quantification of serial pancreas sections. Insulin and glucagon primary antibodies and appropriate secondary fluorescent antibodies are provided. If other staining is desired, the requestor should provide their own antibodies in sufficient amounts to complete their project.
- Frozen Section Insulin Quantitation: The Core provides insulin extraction from frozen pancreatic sections and insulin ELISA assay.
- Quantification of Metabolic Rates in Diabetes-Related Tissues and Cells: The Core supports analysis of cellular respiration and glycolysis and the real-time kinetics of bioenergetic pathways using Seahorse XF Extracellular Flux Analyzers. In addition to providing access to instrumentation (24 and 96 well formats), the core also provides training in application of this platform for the study of islets and β-cells.
Sample Submission
Chargebacks
| Service | Unit of Billing | Charge |
|---|---|---|
| Consultation | Hour | no charge |
| Provision of Established cell lines | Vial | no charge |
| Islet isolation with hand picking | Mouse | $145.38 |
| Human islet quantification and viability | Islet set | $58.15 |
| Hormone Assay by ELISA | Per sample | $8.46 |
| Islet Culture | Islet set | $70.30 |
| Generation of iPSCs or genetically modified cells | Project | Per GESC |
| Static islet incubation | Islet set or mouse | $129.03 |
| Dynamic perifusion testing | Per experiment | $200.00 |
| Paraffin embedded section IF staining per slide | Stain | $9.01 |
| Frozen section Insulin ELISA | Each | $7.56 |
| Frozen section Insulin Extraction & ELISA, technical time | Hour | $58.15 |
| Islet Morphometry from stained slides | Mouse | $255.86 |
| Seahorse XF Extracellular Analyzer | Hour | $40.71 |
| Seahorse TC plates & sensors (24 well or 96 well) | Each | $86.83 |
| ER stress antibodies for IF (BiP, CHOP) | 20 µl each | $34.89 |