Einstein-Mount Sinai Diabetes Research Center

Human Islet and Adenovirus Core (located at Mt. Sinai)

Leadership

Dr. Adolfo Garcia Ocana diabetes research Mt. Sinai

Core Director:Adolfo Garcia-Ocaña, MD 

Dr. Karen Takane diabetes research Mt. Sinai

Operations Manager: Karen Takane, MD 

Location: Icahn School of Medicine at Mount Sinai

Overview

The Human Islet & Adenovirus Core (HIAC) provides key methodologies, technology and infrastructure to support investigators in the use of human islets for research studies for the Einstein-Mount Sinai Diabetes Research Center (ES-DRC). HIAC personnel have extensive expertise in preparation, handling and use of human and rodent islets for research. Given key species-related differences in islet architecture and function, it is of utmost importance to translate the advances and discoveries made in rodent beta cells and cell lines as well as lower species islets to human islet investigation. Conversely, when important advances are made pertaining to insulin secretion by human islets, mechanistic studies are often best performed in isolated mouse or rat islets, or in insulinoma cell lines.

Objectives

  1. To assist new islet investigators in access to human cadaveric and rodent islets and related cell lines for use in investigator-driven studies.
  2. To provide pure populations of live human beta cells using fluorescence-activated cell sorting (FACS).
  3. To provide and/or facilitate the use of specialized reagents, protocols and transduction methods for gene delivery to rodent and human beta cells and islets that will, in turn, enhance investigator-initiated research.
  4. To conduct assays for the determination of glucose- or secretagogue-stimulated beta cell insulin secretion, beta cell differentiation, beta cell proliferation and survival.
  5. To perform specialized surgery and functional analyses using xenograft transplantation of human islets into immune-compromised euglycemic or diabetic mouse models, and mouse syngeneic or allotransplant of islets or cells into euglycemic or diabetic mouse models.
  6. To assist investigators in study design and data interpretation to advance experimental approaches focused on the molecular and physiologic basis of human islet cell function and dysfunction in types 1 and 2 diabetes.
  7.  To train students, postdoctoral fellows, investigators and technical staff in the design and use of molecular, cellular and physiologic approaches in the study of human and rodent islet biology, and pathophysiology. 

Services

  1. Isolation and preparation of human and rodent islets/beta cells and cell lines for investigator-initiated research, which is augmented by the availability of specific islet/beta cell biological assays for the study of beta cell growth, regeneration, survival, immunobiology and function.
  2. Generation of specific viral vectors (adenovirus and lentivirus) for gene delivery of cDNAs and shRNAs of interest to beta cells and other islet cell types. In addition, we will provide viral vectors for transduction of other cell types and tissues under investigation for non-beta cell/islet investigators.

All these services are available to investigators new to diabetes research, as well as to investigators working on diabetes-related projects that can be enriched and extended by the use of the expertise and facilities of this core.

 

Service ES-DRC members Non-ES-DRC members
Training in Human Islet Procurement and Use $30/training episode or hr. $70/training episode or hr.
aIslet culture $40/islet prep $80/islet prep
bIslet cell dispersion and culture $50/islet prep $100/islet prep
cIslet cell proliferation/survival assays (Brdu/Ki67/PHH3 or TUNEL/Caspase3) $100/assay $200/assay
cIn Vitro Islet Function Assay (GSIS in static incubation) $100/assay $200/assay
dIslet Xenotransplant (not incl. mouse purchase, human islets and per diems) $75/mouse transplant $150/mouse transplant
eIslet Xenograft Assessment (assay-dependent) $75-200/assay $150-400/assay
fRodent Islet Isolation (not incl. animal purchase and per diems) $70-100/animal $140-200/animal
gHuman beta cell FACS sort (user provides islets) $500/sort $1000/sort
Adenovirus Library Access: Existing Virus (10µl aliquot/108-109 pfu/µl) $50/virus $100/virus
Custom Adenovirus Prep (100µl aliquot/108-109 pfu/µl) $600/virus $1500/virus
Custom Lentivirus Prep $600/virus $1500/virus
a,bHuman Islet Adenoviral or Lentiviral Transduction $50/Transdux $100/Transdux
cAssessment of Human Islet Adenoviral Efficiency $50/assay $100/assay
Other service Negotiable Negotiable

aCost is for culturing islets in one 6-well plate for one preparation. If more plates are required the discount will apply.

bCost is for dispersing islets in one 24-well plate for one preparation. If more plates are required the discount will apply.

cPrice for a single service, i.e. one in vitro cell proliferation, survival, transfection efficiency or GSIS experiment per triplicate and with three experimental conditions tested.  There is a 20% discount per assay. If additional assays are requested, for example five GSIS experiments the charge is $400.

dHuman islet transplant in one immunodeficient mouse. For additional transplanted mice there is a 20% discount per additional mouse. Weekly blood glucose measurements are included in the price. If mice need to be treated, the following cost applies:
- Tail vein injection: $8/mouse
- Intraperitoneal (ip) injection: $6/mouse
- Subcutaneous (sc) injection: $6/mouse
- Osmotic pump implant: $45/mouse
- Additional personnel time: $25/hr

eHuman islet transplant assessment assays are:
- Islet graft harvest, fixation, embedding, sectioning and staining for insulin in three sections and beta cell area analysis: $100
- Islet graft harvest, fixation, embedding, sectioning and analysis of beta cell proliferation/death: $200
- IPGTT/OGTT, intraperitoneal or oral glucose tolerance test: $75 performed at MSSM
- hyperglycemic clamp and plasma insulin levels see APC and BARC Fee Schedule.

fRodent islet isolation will be performed in mice ($70/mouse) or rats ($100/rat) using histopaque gradient. If islet handpicking is requested after purification, a $10 fee per animal will be charged. Any additional mouse or rat islet isolation will have the 20% discount.

gCost of human beta cell sorting is per preparation and for two treatments, control and experimental. If no treatments are requested, cost will be half.

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