The MRC London Neurodegenerative Diseases Brain Bank:
a resource for neurodegeneration research
Claire Troakes and Safa Al-Sarraj
Institute of Psychiatry, King’s College London, UK
ABSTRACT: Clinically and neuropathologically well-characterised human brain tissue is one of the
most important resources for neuroscience research and is essential in the battle to develop new
strategies and treatments for neurodegeneration. There has been significant research progress in
recent decades and post-mortem tissue has played a major role in enabling advances in diagnosis,
characterisation of pathological features, molecular genetics and bioinformatics.
The MRC London Neurodegenerative Diseases Brain Bank (LNDBB) is one of the largest brain banks
in the UK. Since its establishment in 1989 it has collected over 2000 cases (formalin fixed and frozen
samples). We focus our banking on neurodegenerative diseases, including Alzheimer’s disease,
Dementia with Lewy Bodies, Motor Neurone Disease and Frontotemporal lobe dementia and agematched controls but also house smaller collections, such as psychosis and paediatric disorders, in
order to enhance research in these areas. The LNDBB operates a transparent and open-door policy for
provision of central nervous system tissue to researchers. So far we have completed over 1270 requests
and provided over 10,000 samples to national and international institutions. We are part of the MRC
UK Brain Banks and Brains for Dementia Research networks which aim to encourage and facilitate
both tissue donation and accessibility and use by researchers.
The brain bank also carries out studies into the best methods of preservation of tissue and the
research potential of archival fixed tissue. We are constantly updating our procedures to ensure tissue
is of the best quality for use in current research techniques.
1. Recruitment of cases
a) “Diseased” brains
Brain donations are accepted in the Brain
Bank via two routes:
I. Donor programme: The majority of
donations from Alzheimer’s disease,
MND and movement disorder patients
are registered through cohort studies.
These donations are ideal and provide not
only well-characterised brain tissue but a
fully documented clinical profile,
sometimes with neuro-imaging and
neuro-psychological information. Such
donations are very useful in many
molecular and genetic studies where
knowledge of clinical and pathological
heterogeneity is important.
II. Ad-hoc donation: This has noticeably
increased in the last few years and is a
very useful source of various types of
dementias, other neurological conditions
and “normal” control tissue.
b) Control brains are collected via
- recruiting normal individuals in cohort
studies;
- liaising with the local Transplant
Coordinating centres and Coroners;
- expanding on ad-hoc donation.
Brain tissue available
Neuropathological diagnosis
Stored material 1989 to
2010
Fixed
TOTAL
4. Database information
We have a comprehensive database of information on registered donors and on donated brain tissue, detailing
clinical and pathological diagnosis, post mortem details of fixed and frozen tissue, CSF etc and non-clinical
data such as age, sex and source of donation. Recent donations are also being added to the MRC and BDR
network searchable databases.
5. Environment and links
MRC Centre for Neurodegeneration
The MRC Centre for Neurodegeneration Research at the Institute of Psychiatry aims to understand the
mechanism of neurodegeneration and to translate this into new treatments by carrying out translational research
and facilitating new collaborations. The Brain Bank occupies a key role in this and is a critical resource for the
execution of the centre’s research.
Brains for Dementia Research (BDR)
The Brain Bank is a member of the Brains for Dementia Research network which is funded jointly by the
Alzheimer’s Research Trust and the Alzheimer’s Society. King’s College London acts as the coordinating
centre and the Institute of Psychiatry is one of 6 component Brain Banks. BDR aims to create and maintain a
web based database for all accessible samples in the component brain banks, host training in organ retention,
ethics and governance issues and introduce dedicated Brain Bank research workers who will be responsible for
liaising with the donor and their families at the time of consent, the time of donation and afterwards.
MRC UK Brain Banks Network
The Brain Bank is also a member of the new MRC UK Brain Banks Network. This is an independent and
coordinated national network of existing brain tissue resources. The network will seek to provide operational
efficiency for the benefit of donors, researchers and future patients.
Frozen
1716
Steering Committee
1452
EXAMPLES
Alzheimer’s disease (AD)
459
368
AD-familial
33
14
AD & Cerebral Vascular Disease (CVD)
31
17
AD & DLB
66
58
Autism
4
4
Argryophilic grain disease
8
5
Cerebral vascular disease
67
43
Oxford
12
8
Prion disease (CJD)
27
40
Dementia with Lewy Bodies (DLB)
73
57
Down’s syndrome
4
12
Frontotemporal dementia
49
37
Huntington’s Disease
10
8
Motor Neurone Disease
187
180
Multiple system atrophy
21
15
Parkinson’s Disease
26
18
Progressive supranuclear palsy
27
19
Rett Syndrome
5
5
Schizophrenia
1
23
Normal adult brain
156
Newcastle
7. Tissue Quality Research
Brain banks have large collections of fixed human post-mortem tissue stored as FFPE blocks and as wet
tissue in formalin solution. Recent advances have enabled genetic material to be extracted from this
tissue, however it is important to establish the quality of this DNA/RNA and its suitability for use in the
most current research techniques. The effect of storage in liquid fixative on immunohistochemical
reactivity is also important to investigate. We have therefore recently been studying the research potential
of this archival tissue.
6
1
27
b) Brain tissue dissection and diagnosis
Sets of standard protocols are in use for tissue sampling of fresh
and formalin fixed material. The brain is divided along the midline
in the sagittal plane: one half to be fixed, while the other is freshly
sliced. 50 blocks (1cm x 1cm) are sampled from the slices from
predefined regions, snap frozen and stored at -80C. The formalin
fixed half of the brain is examined by the team of senior
neuropathologists to provide diagnosis according to a
comprehensive protocol which not only reaches a definitive
diagnosis, but also includes details of the load and staging of
pathology and other information important for researchers.
29
1
98
10
38
17
2 3
3
1
28
19
35 39
36
4
44
30
32
33 40
45
41
15
42
7
47
49
DNA extraction was carried out from a range of fixed tissues, of various disease classes, stored for a
range of time (between 0 and 20 years) as blocks or in formalin (a total of 57 cases). The quantity and
quality of DNA extracted from original FFPE blocks (taken at time of autopsy), tissue stored in liquid
fixative, and new blocks processed from the wet tissue was examined. The effect of long-term storage in
formalin on the reactivity of tissue to a number of diagnostic and research-relevant antibodies was
investigated by comparing staining intensity in original and newly processed blocks from the same cases,
using tissue microarray technology.
49 11
48
5
50
48
1
2
Overall, original blocks produced a higher quantity and better quality of DNA than either wet tissue or
newly processed blocks. There was no direct correlation between time of storage in formalin and quality
of DNA. Extracted DNA was of high enough quality to carry out PCR reactions but showed poor results
in a gel electrophoresis run. Newly processed blocks showed a decreased level of reactivity with several
currently used antibodies, including HLA-DP and GFAP.
13
Quality/purity of DNA extracted from Dementia with
Lewy bodies cases. An increase in quality of extracted
DNA was seen in the old blocks compared to new blocks
and wet tissue. DNA quality (A260/280 level) in all cases
combined showed no significant correlation with length of
storage time.
Recent tissue provision
3. Tissue dissemination
a) The Brain Bank has a transparent and open-door
policy for providing services and brain tissue to
requestors from any institution without prejudice,
on the condition that ethical requirements are
satisfied and that a scientifically sound case
underlies the application. The researchers are asked
to complete a request application providing an
abstract of the project, source of funding and other
aspects which is assessed by an approval
committee.
8
1
36
4
23
46
37
Bristol
6. Public Engagement
The MRC London Brain Bank is committed to openness and transparency in its regulatory procedures
and the research it supports. It regularly submits articles to patient support networks and newsletters and
participates in public engagement exercises such as Research Open Days. Lay members and charity reps
sit on the Steering committee.
14
2 26
25
4
43
Brain Net Europe
19 Brain Banks
Cardiff
22
20
London Neurodegenerative
Diseases Brain Bank
1989-2010
MRC UK
Brain Banks Network
130
21
Centre for Neurodegeneration
Research
IOP, KCL
Brains for Dementia Research
Coordinating Centre
Manchester
Corticobasal degeneration
2. Post mortem and tissue collection
a) Post mortem delays
Ideally, we seek to minimise the post mortem delay (time between
death and obtaining tissue for freezing and fixation) to less than 48
hours, wherever possible. Generally we accept donations with post
mortem delay of up to 72 hours (to accommodate deaths that occur
over a weekend or Bank Holiday).
King’s College Hospital
Brain Bank Director:
Dr Safa Al-Sarraj
2006
2007
2008
2009
2010
Total
Total completed
requests
18
40
45
49
57
209
Total samples
distributed
539
1401
2325
2204
3570
10,039
Frozen
267
944
1515
1711
2967
7404
Fixed
272
457
810
493
603
2635
Disease categories requested
A
B
GFAP immunohistochemistry (1:4000) : tissue cores from old
FFPE blocks show a higher intensity of staining than those
taken from newly embedded blocks, suggesting an adverse
effect of prolonged storage in formalin.
Tissue requested 2006-2010
CVD
CBD
DLB
b) We have a wide range of national and
international collaborations and are considered to
be a major resource of brain tissue. We have
responded to over 2,230 requests for tissue since
our establishment in 1989 with 209 completed
requests between 2006 and 2010.
FTLD
Control
HD
MND
MSA
Other
Schiz
PSP
The study shows that DNA can be extracted from FFPE blocks and wet formalin-stored tissue and that this
DNA is of a suitable quality to be used in a number of current research techniques. However the best
quality DNA is obtained from the original blocks, suggesting as many as possible should be taken at
autopsy. The effect of storage in formalin on immuno-reactivity indicates that this variable should be
routinely recorded and included in analysis.
PD
Contact Details The MRC London Brain Bank practises an unbiased policy towards all peer-review tissue request applications; however proof of ethical research approval for the research project may be required. Tissue request application forms can be requested from Dr
Claire Troakes, PO 65, Institute of Psychiatry, King’s College London, De Crespigny Park, London SE5 8AF. Claire.troakes@kcl.ac.uk. All applications are treated confidentially.
Acknowledgements: We thank the Medical Research Council-UK for supporting the Brain Bank. We thank the staff of the Clinical Neuropathology Department, including Dr Istvan Bodi, Dr Andrew King and Dr Tibor Hortobagyi, for advice and technical support. We also
thank Vassiliki Spandoni and Richard Hudspith for technical and administrative help.