AINP 42nd Congress
May 24 - 27th, 2006 Rome Italy
Corso Teorico Pratico
Tissue and Brain Banking
The European Huntington Disease Network
(EHDN)
Speaker:
Ing. Pasquale De Blasio
“Coriell - BioRep” Partnership
Global Biorepository “service providers”
Pasquale De Blasio, CEO
BioRep S.r.l.
Via Fantoli 16/15 20138 Milan Italy
www.biorep.it
e.mail: p.deblasio@biorep.it
The Coriell Institute for Medical Research
For over 50 years the global “Gold Standard” for the storage,
procurement, processing, and distribution of both public and
private biomaterials for scientific research and discovery.
The founder Dr. Lewis Coriell designed breakthrough cell
line storage and processing technologies that enabled his
colleagues, Drs. Salk and Sabin, to develop the polio vaccine.
 For over 30 years Coriell has been the primary provider of cell line repository services
to the National Institutes of Health (NIH), the NCI and 50 countries around the world.
 A sampling of Coriell’s public cell repositories includes collections from National
Cancer Institute, the American Diabetes Association and the National Institute on Aging,
Juvenile Diabetes Foundations International, National Eye Institute, National Institute
for Allergy and Infectious diseases, National Institute for Arthritis and Metabolic Diseases,
National Institute for General Medical Sciences.
 With fully certified ISO 9001:2000 cell culture laboratories, Coriell’s ”Best of Class"
QA/QC processes, forms, policies and procedures have established the highest scientific
standards for cell line, tissue, whole blood and DNA purity, processing and storage.
Coriell-BioRep Locations
 Coriell is located in Camden
N.J. USA, next to Cooper
Health System Hospital in the
Philadelphia try state area.
 Mission
The purpose of the Coriell
Institute for Medical Research
is to discover the causes of
genetic
and
non-genetic
disorders in order to improve
the health of human beings
through the banking of cells
and tissues.
 BioRep is located inside of a New
Science Park located in Milan (Italy)
near Linate Airport and the major
fast carriers sites (DHL, TNT,
Federal Express, etc.).




 BioRep
Biorepository Services
University of Milan, Centre of excellence CISI
Technological Platforms: combinatorial chemistry; HTS
genomics; Proteomics; Microarray technology; Bioinformatics
and Molecular modeling for human, animal and plant
research.
MultiMedica MultiLab
Multimedica Hospital “core lab” with anatomy pathology and
clinical chemistry facilities
Integrated Systems Engineering
Bio-medical instrument design (Tissue Arrayer, western bloth,
etc.)
BioSkills
organization of specialized accredited ECM (Continuous
Education in Medicine) courses for the Biotech industry and
Public institutions.
Coriell-BioRep services
• Coriell Institute has a history of achievement and
excellence in cell culture technology spanning more than
four decades.
• Not only has Coriell consistently been in the forefront of
developing the fundamental technology of cell culture, it
has led the design of electronic catalogues for its
Repositories and the implementation of relational
databases as key parts of its bioinformatics mission.
• Since its founding in 1953, Coriell Institute’s scientists
have pioneered in problem’s solutions in cell culture, cell
characterization and cryopreservation, thus contributing
substantially to the current standard of practice in the
field.
Services
• Coriell and BioRep offer a range of services to
public and private research institutions and are
committed to a flexible and innovative approach
in order to meet individuals particular needs:
–
–
–
–
–
–
–
cell culture
molecular biology
cytogenetics
research kits and reagents
cell repository
tissue bank
tissue microarray
Coriell - BioRep Quality Assurance programs
•
•
•
•
•
•
Quality Management
ISO9001:2000 Certification (in progress)
Facility Services GLP/GMP compliant
Testing and Pre-qualifying Reagents
Redundancy of Processing
Redundancy of Storage
– Storage in the main facilities (Milan)
– Back-up facilities (Monza or USA at Coriell)
• Redundancy in the Database
• Cell Line and Tissues Quality Control
• DNA/RNA Quality Control
Coriell –BioRep information System
Coriell - BioRep
BioRep Management System
•
•
•
•
•
Biorep Catalog & Order System
Biorep ERP
DHL tracking system
Coriell’s Queue
Third party system already interconnected:
– EHDN system, Germany
– Coriell, USA
Central Vs. Local BioBanking
Virtual Network
- Local Storage
• Advantages
– Direct control of
specimens
– Immediate availability
locally
– Small Bio-Bank facility
– Easy management of
local research group
- Central Network
• Disadvantages
– Heterogeneity of:
• SOP’s
• IT infrastructure
– Quality of the Bio-Banking
facility and organization
– Difficult to share
specimens
– Difficult to manage multicentres projects
– Difficult to expand
Central Vs. Local BioBanking
Virtual Network
- Central Storage
• Advantages
– Central Coordination of the
projects
– Same:
• SOP’s
• IT infrastructure
• Logistics
– Large and organised BioBanking facility
– High quality and Safety
standards
– Room for expansion
– Controlled access to the biospecimens stored
- Central Network
• Disadvantages
– The Network needs a
formal organization
– Long-Term Founding of
the Central Bio-Banking
facility
– Access roles to the biospecimens
Central Vs. Local BioBanking
High Q supports HD Clinical Research through the
Huntington Project
Cohort
Registry
Huntington Project
Euro HD Network
USA
Europe
Central Biorepositories
Coriell Institute for
Research, NJ-USA
BioRep
Medical
Milan, Italy
USA - Canada
14 European Countries
90 Centres involved
Collection sites
104 Centres involved
Collection sites
Current collection kit
• Contains all items required to package and send
samples
• Packaging and shipping cost per subject approx 40
euros ($33) (exc. labour and overheads)
Advantages:
• Comes flat-pack reducing shipping costs
• Relatively cheap & easy to use
Disadvantages:
• Limited protection for samples
• No temperature control - when exposed
to 37°C external heat, internal temp
raised by 10°C (22 to 32°C) in approx 1hr.
Dual compartment box
• One compartment at ambient temperature with 2 gel
packs, and one at approx 4°C using 2 frozen gel packs
• Packaging and shipping cost per subject approx 94
euros ($78) (exc. labour and overheads)
Advantages:
• Maintains temp on cold side for 30hrs
with external temp of 20°C and 12hrs with
external temp of 37°C
• Greater protection for samples
Disadvantages:
• High shipping costs
• Bulky
• Packaging procedure more complex
EHDN Registry Repository
May 2006
REGISTRY REPOSITORY
• Preparation and disposition of DNA from whole
blood for CAG repeat measurement
– Isolation of DNA from the whole blood contained in one
ACD tube using a modification of the Miller salting out
procedure [Miller, S.A., Dykes, D.D., and Polesky, H.F.
A simple salting out procedure for extracting DNA from
human nucleated cells. Nucleic Acid Res. 16: 1215
(1988)].
– Prepare one 25 microgram aliquot of the DNA and
dividing the remaining DNA between two additional
aliquots (for a total of three aliquots per subject).
– Store the 25 microgram DNA aliquot
– Measure CAG repeat number.
REGISTRY REPOSITORY
• Isolation of lymphocytes and establishment of
lymphoblastoid cell lines
– Isolate lymphocytes from the whole blood contained in
one ACD tube.
– Divide the lymphocytes into four aliquots.
– Use the aliquots of lymphocytes as necessary to
establish a lymphoblastoid cell line from the
lymphocytes.
– Check the cell line for viability, identity, and freedom from
contamination by bacteria, fungi, mycoplasma, and other
organism and cell lines.
– Prepare 10-20 aliquots of the cell lines.
– Store lymphoblastoid cells lines in liquid nitrogen
– Store the remaining aliquots of lymphocytes
REGISTRY REPOSITORY
• Preparation of DNA from lymphoblastoid
cell lines
– Expand each lymphoblastoid cell line to produce
approximately 200 micrograms of high quality, high
molecular weight (>50 kb) DNA.
– DNA will be isolated from the lymphoblastoid cells using
a modification of the Miller salting out procedure [Miller,
S.A., Dykes, D.D., and Polesky, H.F. A simple salting out
procedure for extracting DNA from human nucleated
cells. Nucleic Acid Res. 16: 1215 (1988)].
– Perform quality control studies to estimate of quality and
integrity and verify identify.
– Prepare 25 microgram aliquots of the DNA
– Store DNA as set forth below.
REGISTRY REPOSITORY
• Preparation of plasma
– Prepare plasma from the whole blood contained in
one EDTA tube.
– Prepare 0.5 ml aliquots (20 max) of the plasma.
– Store plasma as set forth below.
• Preparation of urine
– For subjects participating in Registry, prepare up to
six 4 ml aliquots of urine.
– Store urine as set forth below.
EHDN – SAMPLES RECEIVED
Registry repository contains 69 subject samples
EHDN REGISTRY REPOSITORY
NO. SAMPLES
RECEIVED
30
25
Madrid
20
Lisbon
15
10
Barcelona
Ulm
5
Graz
0
Nov-05 Dec-05 Jan-06 Feb-06 Mar-06 Apr-06
MONTH
FUTURE EHDN CENTRES
Approx 30 additional centres in next 3 months
Region / Countries
No. Centres
Locations
Scandanavia
4
Oslo, Bergen, Turkku, Lund
Benelux
1
Lieden
Denmark
1
Copenhagen
Italy
1+4
Milan + 4 sites
Spain & Portugal
2
Valencia, Granada
UK
6
Cardiff & 5 others
Germany
10
To be confirmed
Switzerland
1
Bern
DHL DELIVERY TIMES
• Av. time from collection to delivery = 25hrs
• Longest delivery time = 171hrs (Xmas)
• Shortest delivery time = 15hrs
• 81% shipments received within 24hrs
• 96% shipments received within 48hrs
BioRep Cell Culture Laboratorie
Coriell-BioRep Cell Culture Services
• Establishment of cell lines
– EBV transformed lymphoblast cultures from human blood
– fibroblasts, endothelial, smooth muscle and epithelial cell cultures
from appropriate tissue biopsies
•
Cell banking of public, private, or specialty collections
– cryopreservation of primary or transformed cell lines
– cryopreservation of peripheral blood mononuclear cells (buffy coats)
and selected tissue biopsy material
– maintenance of seed and distribution stocks
– expansion of cell lines for molecular and cytogenetic analysis
• Mycoplasma testing
– detection by PCR or microbiological assays
– identification of mycoplasma species by immunological or PCR
methods
• Remote Fail-Safe Storage
– cell lines
– DNA
– blood products and other biomaterials
BioRep Molecular Biology Laboratories
BioRep Molecular Biology Laboratories
BioRep Molecular Biology Services
• DNA Isolation
– microgram to milligram quantities of high molecular weight DNA isolated
from whole blood, cell lines, and tissue biopsies suitable for molecular
biology applications
• RNA Isolation
– polyadenylated (mRNA) and total RNA from a variety of cell lines and
tissues verified for quality by RT-PCR
• Genetic Mapping
– mapping the chromosome location with genetic probes
• Whole Genome Amplification (WGA)
– high fidelity amplification of DNA from limited amounts of fresh or frozen
genomic materials (e.g. from mouthwash samples)
• Gender Analysis
– gender analysis based on the fluorescent detection of sex chromosome
specific alleles
• Verification of family pedigrees
– genotyping with highly polymorphic microsatellite markers
• Promoter Methilation analysis
– level of methilation in CpG island
• SNPs analysis via DHPL method
Coriell Cytogenetics Services
• Coriell's cytogenetic research group has three decades of experience in
chromosome analysis and, using the most advanced techniques, is
fully equipped for bright field and fluorescence microscopy. Digital
imaging and analysis are performed using the Applied Imaging
Cytovision Ultra and the Vysis Smart Capture Imaging System.
•
•
•
•
•
Karyotyping (Human)
– standard analysis of metaphase chromosome using G-banding technique
– high resolution analysis of prometaphase chromosomes using G-banding and other banding
procedures as required
Karyotyping (Mammalian)
– standard analysis of metaphase chromosomes from a wide variety of mammalian species,
including but not rrestricted to mouse, dog, pig, as well as mouse stem cells
– standard metaphase chromosome analysis of various primate species
FISH Analysis
– mapping
– localization of chromosome break points
– estimation of gene copy number in interphase
– determination of interphase zygosity of transgenic mice
Comparative Genomic Hybridization Analysis (CGH)
– assessment of chromosome loss, gain, or amplification in a sample of genomic DNA by
competitive hybridization of two genomes against a target cell
Chromosome Aberration Analysis and Micronucleus Assays
– evaluation of genomic damage by a variety of assays to provide genotoxic end points
Coriell - BioRep Cell and Tissues repositories
• The CORIELL and BioRep REPOSITORIES
provide essential research reagents to the
scientific
community
by
establishing,
verifying, maintaining, and distributing cell
cultures and DNA derived from cell cultures
and
tissues.
These
collections
are
extensively utilized by research scientists
around the world.
•
•
•
•
•
Bar-coding and tracking system for all bio-specimen
vials
Storage in:
– Liquid nitrogen at -196°C or vapour phase
(lower than -150°C)
– Mechanical (standard and automated) freezers
at -80°C (or - 150°C)
– Additional storage at +4°C and room
temperature
– Storage for customer-owned freezers and
their contents
PC/PLC based “proprietary” state-of the-art
alarms and monitoring system
with continuous
24x7 operations.
100% back-up redundancy / off-site sample storage
of all cell and tissues
Remote storage
Prodotti BioRep
BioRep Sistema Vendita
di Monitoraggio
e Controllo
• Monitoring and Control System
Characteristics:
• Ethernet, RS232, e Wirless
connections
• 24/7 - 365 days/years
continuous monitoring and
recording
• Alarm system based on 3 levels
of alarms (visual, sound, SMS)
• Web visualization
(www.biorep.it/servizio)
accessible with Password
• Certification reports (charts,
tables, etc.)
BioRep Management System
•
•
•
•
•
Biorep Catalog & Order System
Biorep ERP
DHL tracking system
Coriell’s Queue
Third party system already interconnected:
– EHDN system, Germany
– Coriell, USA
Biorepository Management System
BioRep web portal
• Access through Biorep web- portal (www.biorep.it)
• User self registration
• Subscribed User
– Purchase services & biomaterial from catalog
BioRep Tissue Microarray Services
• preparation of custom tissue arrays and distribution of tissue array
slides from specific needs
• The new computer controlled tissue arrayer is developed in
collaboration with ISE and:
–
–
–
–
–
NIH Tissue Array Research laboratory (Dr. Stephen M. Hewith)
Erasmus University of Rotterdam NL (Dr. Peter Riegman)
CNIO of Madrid (Dr. Manuel Morente)
National Cancer Institute of Milan (Dr. Paolo Carbone)
Multimedica MultiLab (Dr. Massimo Barberis)
The Tissue MicroArrayer
Specifications

High resolution optics

Automated
and
needles positioning

Automated punching depth regulation

Up-to-6 block automated tray

Dedicated software to assist the user
in all operating phases, from the TMA
template definition to the final
reporting
computer-guided
Galileo TMA CK3500
-
Integrated Systems Engineering srl, Milan, Italy
The Tissue MicroArray (TMA) technology
 Simultaneous histological analysis of
hundreds of samples using the
minimum amount of reagents.
 Use of relatively small areas of tissue
(down to 0.6 mm in diameter)
 Cost effective technique
 Saving
of
specimens
precious
biological
TMA Design
 Software assisted
 Performed remotely by the arrayer
TMA_Adenoma
TMA Design
To optimise the tissue core placement
for subsequent analysis
TMA_Adenocarcinoma2
TMA_Adenocarcinoma1
Lesion
Normal Tissue
(near lesion)
Case X
Case Y
Lesion
Normal Tissue
Control Tissue
Normal Tissue
(resection margin)
TMA construction
The sampling procedure
TMA analysis
SEL1L anti-body expression evaluation
Case Number
1
2
3
4
5
6
7
8
9
10
11
L
L
L
L
L
NM NM NM NM NM NM NM NM NM
SN 1 SN 2 SN 3 SN 4 SN 5 SN 1 SN 2 SN 3 SN 4 SN 5 SNv1 SNv2 SNv3 SNv4
0
0
1
0
0
0
ND ND
0
ND ND
0
ND ND
0
ND
0
0
0
SEL1L expression intensity
0
0
0
ND
0
ND
3
Negative
2
ND
3
0
0
1
2
1
1
0 0
2
1
1
1
1
2
2
1
1
1
1,2,32 Positive
0
0
0
0
0
2
2
2
1
1 cells
2 (Reference)
3+ 1 Plasma
2
2
2
1
1
ND
2
2
0
3
1
1
1
ND
0
2
2
ND
2
2
ND
ND
0
0
ND
0
0
0
0
1
0
0
0
0
0
0
1
0
Adenocarcinomas
Adenomas
Not sampled
ND Not Defined
INTERNATIONAL GUIDELINES FOR
BIOBANKS
Milano, 16 giugno 2006
Location:
Sala Conferenze
BioRep srl
Via Fantoli 16/15
Milan (MI) - Italy
Thank you for your collaboration and kindness
from the BioRep team