Chicago Cell Biology Core

Overview

The Cell Biology Core provides services and hands-on training in the isolation and functional characterization of pancreatic islets from normal and diabetic humans and mice. In addition, it maintains a repository of insulinoma cell lines for distribution. The primary emphasis of the Core is to facilitate studies of primary islet cells and it has developed many unique tools and techniques for carrying such studies including novel animals models, biophysical methods and a library of adenovirus-based expression constructs for studying beta-cell function. The Cell Biology Core provides state-of-the-art technology and know-how to understanding the beta cell in health and disease.

Objectives

The overall objective of this Core is to provide high quality islet and insulin–secreting cells to investigators together with basic and advanced methods to evaluate islet function. Specific objectives are:

• Advice, service and training in the isolation of pancreatic islets from normal and diabetic rodents and humans
• Provide insulinoma and other endocrine cell lines and advice on their use
• Advice, service and training in the characterization of rodent and human pancreatic islets and beta cells
  • Proinsulin biosynthesis, processing and insulin secretion
  • Biophysical methods for studying islet and beta-cell function (e.g. calcium imaging, electrophysiology, total internal reflection fluorescent microscopy)
  • Biochemical methods for studying beta-cell function (e.g. phosphorylation, proliferation, apoptosis)
  • Immunohistochemical analysis of pancreatic islets
• Advice, service and training on the use of adenovirus-based expression constructs to study protein function in beta cells and other cell types in vitro and in vivo

Services

Provide rat, mouse and human islets from normal and diabetic sources.

The staff obtains the appropriate animals and using approved protocols, prepares islets and initiates cultures or other special handling as requested. They also supervise or teach the assays of hormone secretion that are part of the quality control protocols.

Maintain insulinomas and insulinoma cell lines for distribution.

Dr. Steiner’s laboratory maintains a large number of insulinoma, neuroendocrine, and other relevant cell lines. The laboratory directly confers with Investigators and provides flasks of cells or frozen cells at particular passage numbers as requested, and from time to time test the viability and quality of specific lots. The laboratory maintains logs of cell lines available, storage and growth conditions, passage number, and quality control aspects, such as glucose-stimulated insulin secretion and mycoplasma monitoring. A partial listing of available material includes HIT, RINm5f, BTC3, BTC6, INS-1 (several kinds), MIN-6, HEK, COS-7, CHO, HeLa and SF9. A large number of mouse, rat, hamster and human insulinoma samples are also available.

Provide training in isolation and culture of pancreatic islets from mice, rats and humans.

A major function of the laboratory is to provide training in islet isolation, culture and biology. Interested faculty, fellows, and students are provided with hands-on training from experienced technicians so that they can gain first hand knowledge of these important skills.

Provide service and training in methods for studying insulin synthesis and secretion using a variety of methods.

Insulin biosynthesis has been a main focus of Dr. Steiner’s laboratory. A variety of approaches have been provided for studying islet hormone biosynthesis in great detail. Insulin secretion can be studied in a variety of ways. Both static and dynamic approaches to insulin secretion measurements are available. Direct and indirect, biophysical measures are also provided. These studies are carried out in close coordination with the requesting investigator.

Provide service and training in methods for immunohistochemical analysis of islets and insulin-secreting cells.

Investigators desiring these services consult with the Director and determine the appropriate reagents and conditions for particular studies. Histochemical studies are coordinated with the assistance of the Pathology section of the Animal Resource Center at the University of Chicago, the Electron Microscopy Core, and the Digital Imaging Core. Digital images are prepared for publication. Archival data storage of key images, reconstructions, and movie files are also maintained.

Provide service and training in calcium imaging and other biophysical methods for studying islet and beta-cell physiology.

Investigators have available a number of techniques, including optical measurement of NAD(P)H, Rh123, TMRE, fura-2, fluo-3, MgGreen, FuraRed, and MgFura. Also included is detection of FRET (fluorescence resonance energy transfer) substrates and use of targeted GFP-based fusion proteins. The emphasis of the Core is on specialized live-cell techniques and dedicated islet perfusion systems. The Core also provides an interface to appropriate Divisional Cores, such as the Digital Imaging Core and Electron Microscopy Core of the Division of Biological Sciences. Through collaboration with Dr. Norbert Scherer in the Department of Chemistry, multiphoton imaging analysis is also available.