Einstein-Mount Sinai Diabetes Research Center

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

Contact Info

Hours: 9:00 AM - 5:00 PM

Locations: Icahn School of Medicine at Mount Sinai
Berg Laboratory Building Floor 5th Floor Room AB5-10
1428 Madison Avenue, New York, NY 10029

Phone: Adolfo Garcia-Ocana, PhD, 212.241.9793

Email: adolfo.garcia-ocana@mssm.edu 

 

 
 

Leadership

Dr. Adolfo Garcia Ocana diabetes research Mt. Sinai

Core Director: Adolfo Garcia-Ocaña, Ph.D. 

 
Dr. Peng Wang

Operations Manager: Peng Wang, Ph.D. 

 

Location: Icahn School of Medicine at Mount Sinai

Overview

The first mission of the HIAC is to provide key advice, methods, technology and infrastructure to assist investigators in the use of human islets for research, with the goal of furthering understanding of normal and pathophysiologic islet cell growth and function. Although rodents unequivocally continue to provide new important insights into islet biology and pathobiology, it is of utmost importance to translate the advances and discoveries made in rodent beta cells and cell lines to human islet investigation and therapies. Conversely, when important advances are made in human islet cells, mechanistic studies are often best performed in isolated mouse or rat islets, or in insulinoma cell lines. HIAC personnel have extensive expertise in the preparation, methodology, handling and use of both human and rodent islets, and all relevant insulinoma and alpha cell lines. Thus, while the focus is on human islets, the HIAC also has provided, and will continue to provide, isolation of rodent islets, access to rodent insulinoma cell lines, and specific experimental analyses such as assays for insulin secretion, islet bioenergetics, beta cell proliferation, mass and survival, as well as human islet transplantation into immunodeficient mice, with subsequent functional analyses.The second mission of the HIAC is to generate and make available to the ES-DRC community reagents and tools including adenovirus or lentivirus viral vectors for gene delivery of cDNAs and shRNAs of interest to beta cells and other islet cell types to study beta cell regeneration, differentiation, survival and function. These reagents are also available for use in the transduction of non-islet cell types and tissues.

Objectives

  1. To assist new islet investigators in accessing human and rodent islets and related cell lines for use in investigator-driven studies. 
  2. To train students, postdoctoral fellows, investigators and technical staff in the design and use of molecular, cellular and physiologic approaches to human and rodent islet biology and pathophysiology. 
  3. To provide pure populations of live human beta cells using fluorescence-activated cell sorting (FACS) and/or to facilitate the use of specialized protocols, adenovirus/lentivirus and transduction methods for gene delivery to rodent and human beta cells and islets that enhance investigator-initiated research.  
  4. To conduct specialized assays for the determination of insulin secretion, islet bioenergetics and beta cell differentiation, proliferation, survival and mass in vitro and in vivo using syngeneic, allotransplant or xenotransplant of rodent/human islets into immunocompromised euglycemic or diabetic mouse models.  
  5. To assist investigators with 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.  

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 cell number (Beads+Flow cytometer)  $300/assay $600/assay
cIn Vitro Islet Function Assay (GSIS in static incubation/GSIS by perifusion/Mitochondrial Function, Seahorse Analysis) $100/assay
$300/assay
$300/assay 
$200/assay
$600/assay
$600/assay 
dIslet Xenotransplant (not incl. mouse purchase, human islets and per diems) $75/mouse transplant $150/mouse transplant
eIslet Xenograft Assessment (assay-dependent) $75-300/assay $150-600/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
hAdenovirus Library Access: Existing Virus (10µl aliquot/108-109 pfu/µl) see below $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, number,survival, transfection efficiency, GSIS or mitochondrial function analysis experiment 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/$1300.

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.

h

CMV Promoter 

shRNA  Tet-Inducble 

GFP 

PKB/Akt-WT 

hCdk4 

hp14arf 

 p14    SMAD1  

eGFP 

PKB/Akt-CA 

hCdk6 

hp16 

 p16     SMAD2  

CMV TTA (tet transactivator) 

LacZ 

PKB/Akt-KD 

hCyclinD1 

hp18 

 p18     SMAD3  

TRE-hCdk4 

LacZ-GFP 

PI3K 

hCyclinD2 

hp19 

 p19     SMAD4  

TRE-hCdk6 

PDGFRA 

PTEN 

hCyclinD3 

hp21 

 p21   SMAD5  

TRE-CyclinD1 

PDGFRB 

STAT5 

hCdk1 

hp27 

p27  

SMAD7 

TRE-CyclinD3 

cMet 

JAK2 

hCdk2 

hp57 

p57  

SMAD8 

TRE-cMyc 

PRLR 

PKCζ-WT 

hCyclinA 

hp53 

pRb  

KDM5B 

RIP1-Driven 

PTHrP 

PKCζ-CA 

hCyclinE 

4mtD3cpv 

p107  

KDM6A 

RIP-LacZ 

HGF 

PKCζ-KD 

hBmi1 

long TmfSF4-mcherry 

p130  

KDM6B 

RIP-ZsGreen 

PL 

NFaT2 

hEzh2 

Bach2 

GC  

lats1 

RIP-mCherry 

Betacellulin 

mCherry-NFATc3 

hcMyc 

Tag-BFP 

DYRK1A  

lats2 

RIP-TTA 

VEGF 

ICUE3 

hcMyc-S62A 

BNC2 

DYRK1B  

Dlk1 

RIP-Medial Go-D1 

Pericam 

D4ER 

hcMyc-T58A 

ZRANB 

DYRK2  

HDAC6 

RIP-Trans Go-D1 

GC 

Dlk1 

B-catenin-y33 

GAS7 

DYRK3  

HDAC8 

RIP-VSVG-GFP 

TagBFP 

DYRK1A 

ISL1 

VIVT 

DYRK4  

LacZ 

RIP-GJD2 

Cre 

DYRK1B 

Mybl2 

ChREBP 

GSK3ß  

p300 

IRX1-cherry-rip1-zsgreen 

GSK3α 

DYRK2 

Kdr 

Glucokinase 

GLP-1R  

Creb3 

Tm4sf4-cherry-rip1-zsgreen 

FOXM1 

DYRK3 

SST2 mcherry 

Keap1 

CREBBP  

CLK1 

RIP-CyclinD1 

SMAD6 

DYRK4 

SST5 mcherry 

Nrf2 

Scramble  

CLK2 

RIP-Cdk4 

Sp1 

  

  

  

Men1  

CLK4