The Core provides access to diabetes research-related cell lines:

Cell Line Name	Description
β-TC	Mouse beta cell
ISL-1	Rat insulinoma
266-6	Mouse pancreatic epithelial
Min6	Mouse Insulinoma (SV40 T-antigen induced)
AR42J	Rat pancreatic exocrine cell line
5KC_12-4.1	T cell transductant cells expressing mouse insulin-reactive TCR
5KC_GSE.20D11	T cell transductant cells expressing human insulin-reactive TCR
K562_HLA-DQ8	K562 cell lines expressing HLA-DQ8  (DQA1*03:01-DQB1*03:02)
K562_HLA-DR4	K562 cell lines expressing HLA-DR4 (DRA1*01:01-DRB1*04:01)
K562_HLA-A2	K562 cell lines expressing HLA-A2 (A*02:01)
K562_HLA-A24	K562 cell lines expressing HLA-A2 (A*24:02)
Priess	EBV-transformed B cell line (HLA-A2-DR4-DQ8)

 

Contact:

Maki Nakayama, PhD
1775 Aurora Ct., Aurora CO 80045
Office: 303-724-4076
[email protected] 

Researchers have access to a range of histology instruments. Training for instrument use is provided upon request. We also provide technical assistance and consultation for all aspects of diabetes related histological analysis. Commonly used antibodies and reagents for diabetes research, such as insulin antibodies, are available, along with optimized staining protocols. DRC members requiring paraffin embedding services are referred to the Pathology Shared Resource.

 

1. Instrumentation(s):

• Microm HM 325 Cryostat
• Microm HM505E Cryostat
• Leica RM2155 Microtome
• Olympus CX31 Light Microscope
• Perkin Elmer Vectra 3.0 (in partnership with the University of Colorado Human Immune Monitoring Shared Resource)

 

2. Access to Instrumentation

Researchers have access to a range of histology instruments. Training for instrument use is provided upon request.

• Tissue processor: Miles Scientific Tissue-Tek VIP
• Automated embedding system: Leica
• Microtomes: RMC
• Cryostats: Microm 325
• High-throughput quantitative pathological and immunohistochemical analysis: Perkin Elmer Vectra 3.0 (in partnership with the University of Colorado Human Immune Monitoring Shared Resource)
  

 

3. Technical Assistance, Training, and Consultation

We provide technical assistance and consultation for all aspects of histological analysis. Commonly used antibodies and reagents for diabetes research, such as insulin antibodies, are available, along with optimized staining protocols.

 

For information on histology services, contact:
Lori Sussel, PhD
Barbara Davis Center
1775 Aurora Ct. Aurora, CO 80045
[email protected]

The DRC Tissue Procurement Core coordinates access to human pancreatic islets and other human tissue specimens for registered investigators, leveraging partnerships with major islet isolation centers and biorepositories.

Human Islet Distribution

1. Source & Partnerships:

Human islets are sourced through the University of Alberta Edmonton Islet Isolation and Distribution Program and the Alberta Diabetes Institute. The Core arranges distribution for registered DRC investigators and maintains a Master Material Transfer Agreement (MTA) to facilitate access.

2. Eligibility & Ordering:
Researchers must provide:

• Institutional Biosafety Committee (IBC) approval
• A brief project description
• A signed MTA
  
  

3. Cost & Availability:

• Human islets are available at $0.10 CAD/IEQ (approx. $75 USD per 1,000 IEQ) plus a $50 USD shipping fee.
• Islets are typically offered every one to two weeks.
• Orders are placed through the Core and delivered for distribution.
  
  

Other Human Tissue Specimens

• The Core arranges access to fresh, viable human tissue specimens in partnership with the University of Colorado Biorepository Core Facility (Department of Pathology).
• Specimens are provided with de-identified clinical data to support research needs.

Contact:
Scott Beard 
Barbara Davis Center (Room 4209, 4th Floor)
1775 Aurora Ct. Aurora, CO 80045
303-724-6832 (office)
[email protected]

The CU DRC Tissue Procurement Core offers comprehensive services for the isolation of functional pancreatic islets and other tissues from rodents. These services are designed to support a wide range of research needs and include:

• Rodent Islet & Tissue Isolation Service: Isolation of pancreatic islets from mice, with the option to collect other organs (such as spleen, pancreatic lymph nodes, and thymus) upon request.
• Training for Rodent Islet Isolation: Hands-on training for researchers in islet isolation techniques.
• Reagents for Rodent Islet Isolation: Provision of optimized reagents, such as collagenase, specifically formulated for efficient islet isolation.

 

1. Islet Harvesting Procedure

• Donor Mice: Clients provide the donor mice.
• Euthanasia and Enzyme Injection: Donor mice are humanely euthanized. A collagenase/neutral protease cocktail is injected via the common bile duct to inflate the pancreas.
• Islet Isolation: The enzyme-inflated pancreas is removed and incubated at 37°C. Islets are then purified using a density-gradient and hand-picked under a microscope.
• Reporting: Clients may request islet yield to be reported as a simple islet count or as islet equivalents (IEQs; 1 IEQ = an islet roughly 150mm square).

 

2. Islet and Tissue Transplantation

• Transplantation Service: Isolated islets (scored as IEQs) can be transplanted under the kidney capsule of anesthetized recipient mice (also provided by the client).
• Other Tissues: This transplantation service is also available for other tissue types.
• Animal Housing: Arrangements can be made to house transplanted animals in the BDC Animal Facility.

 

3. Animal Protocol Requirements

• IACUC Approval: Researchers must have a current IACUC-approved animal protocol.
• Protocol Amendment: The Core staff member must be added to your protocol, and a section on islet harvest/transplantation procedures must be included.
• Documentation: The Core will provide the necessary personnel qualification forms (PQF) and protocol narratives to facilitate compliance.
  

 

4. Service Request and Scheduling

• Request Process: Services are requested through iLabs (Barbara Davis Center Bioresource Core).
• Scheduling: Actual scheduling of isolations must be arranged by directly contacting the Core via phone or email.

 

Contact:

Scott Beard 
Barbara Davis Center (Room 4209, 4th Floor)
1775 Aurora Ct. Aurora, CO 80045
303-724-6832 (office)
[email protected]

What is Islet Perifusion?

Perifusion enables assessment of dynamic hormone secretion by primary (e.g., rodent, human) or stem cell-derived islets. Islets are loaded into chambers and connected to a pump and collection system housed in an environmentally controlled enclosure for the application of stimulation solutions and collection of eluate fractions for hormone quantification. Fractions are collected every minute for high temporal resolution of hormone secretion. A manifold controls uptake of up to 8 different stimulation solutions applied to islet chambers in an automated, user-programmable manner to accommodate diverse applications. Common applications include mechanistic studies of experimental or disease models and advanced benchmarking of stem cell-derived islets.


Protocol requirements

Primary or stem cell-derived islets are provided by the end user. Alternatively, islets can be isolated from mice provided by the end user through the TPP Core on a fee-for-service basis, provided that their IACUC-approved animal protocol covers islet isolations and that Mr. Scott Beard has been added as personnel on the animal protocol.

Costs

• $150 per perifusion run and an additional $50 per chamber/replicate.
• DRC members receive a 15% discount.

Service request and scheduling process

• Trained users can request perifusion runs through iLabs (Barbara Davis Center Bioresource Core).
• To request a run, go to Request Services -> DGSIS via Perifusion -> DGSIS via Perifusion User Run – per run -> initiate request.
• Then to indicate number of chambers, go to Request Services -> DGSIS via Perifusion -> DGSIS via Perifusion User Run – per sample -> initiate request.
• Reserve your time by accessing the calendar: go to Schedule Equipment -> Islet Perifusion System -> View Schedule.
• Each end user must be trained directly by the Wortham lab. To schedule training, email Matt Wortham [email protected]. 
  

Samples generated

At the end of the experiment, you’ll have several 96 well plates containing collected fractions that can be used for hormone quantification (e.g., by ELISA) to be performed by the end user. Depending on user preferences, secreted hormone levels can be normalized to hormone or DNA content of islet lysates (processed after the perifusion experiment). 

Contact Information
Matt Wortham
[email protected]
(303) 724-4761