© BrainNet Europe II
BrainNet Europe is a "Network of Excellence" funded by the European Commission in the 6th Framework Program "Life Science" (LSHM-CT-2004-503039). It consists of 19 established brain banks across Europe and is coordinated by the Centre for Neuropathology and Prion Research Ludwig-Maximilians-University Munich, Germany.
Participant 10: Netherlands Brain Bank
The Netherlands Brain Bank
The Netherlands Brain Bank (NBB) is a department of the Netherlands Institute for Neuroscience, an institute of the Royal Netherlands Academy for Arts and Sciences (KNAW). The NBB has a close cooperation with the VUmc (the medical centre of the Vrije Universiteit). The VUmc takes care of the autopsies and diagnosis. The NBB (NBB) was established in 1985 to support neurobiological and medical research by supplying clinically and neuropathologically well-characterized post-mortem specimens from patients who suffered from neurological and psychiatric disorders.
Due to the large variability of the collected material, there are many drawbacks in the use of post-mortem specimens. Therefore, brain banks should put special emphasis to the development of rapid autopsy systems, i.e. as practiced by the NBB (Ravid and Swaab 1993) and guarantee the quality of the specimens by proper matching for the various ante and post-mortem factors and by measuring the pH (Ravid et al., 1992).
The NBB has three unique features:
(1) The bank has established a national donor-program, consisting of approximately 1800 registered donors. All donors and their next of kin are requested to sign an informed consent, when joining the program. In addition to the donation of brain, spinal cord and body fluids for scientific research, the donors are able to donate some tissues for transplantation and whole body for the anatomy.
(2) Human brain tissue is obtained by means of rapid autopsies with a very short post-mortem delay, ranging between two to eight hours.
(3) A fresh brain dissection procedure is used. The latter is a difficult regime to establish, requiring qualified staff at inconvenient times, but is a prerequisite for an increasing range of technical procedures, such as cell culture, and for neurochemical, immunocytochemical and metabolic procedures. At autopsy, the pH of the cerebrospinal fluid (CSF) is determined as a measure for agonal state, which is of importance for, e.g., the yield of mRNA.
Once the brain has been removed it is macroscopically examined and immediately dissected following a standard protocol into 80 different structures. In addition to brain samples we also collect ventricular CSF in each rapid autopsy for measuring the pH of the brain and for subsequent investigation of biological markers and other compounds present in the CSF in various disorders. After centrifugation, the CSF is aliquotted into 1ml small tubes and then slowly frozen and stored in - 80 C. The freezers of the brain bank have a liquid nitrogen backup system and are carefully monitored to assure continuous temperature maintenance. Automatic signal alarm systems are used to maintain the right temperatures.
The Brain Bank has at its disposal a growing collection of body fluids (CSF, blood and plasma) obtained from a wide range of disorders as well as a large series of formalin fixed brain tissue. Donor tissues that are acquired for storage include also spinal cord, cerebral arteries, olfactory bulb, pituitary and optic nerves.
A standard number of approximately 16 samples are fixed in formalin, further processed, and stained by various histological or immunocytochemical methods for diagnostic purposes. To prevent contamination of the dissected tissue and to make all specimens suitable for isolation of high yield m-RNA we use semi-septic methods including sterilization of instruments and working surfaces and use of protective gloves for personnel during tissue handling.
The NBB collects a comprehensive medical history from each donor after autopsy and we try to include as much relevant information available on chronic and acute diseases, their course and medication used, alcohol and drug abuse and smoking habits. All medical records are secured with informed consent of the donor or next of kin requested in advance together with the informed consent for performing the autopsy.
Shipped samples are always accompanied by a D-base printout specifying the major items such as age, gender, brain weight, post-mortem delay, pH (agonal state), clock time of death, types of tissue and anatomical structures acquired from the donor and a summary of each donor's medical history.
The final diagnosis is established after an extensive neuropathological examination and relates the findings to the clinical diagnosis. The definite diagnosis is sent to the patient's physician as well as to the various research groups. The neuropathology reports are produced in a highly standardized format, which on its turn increases significantly the utility of such reports for international investigators. The conclusions and summary of the neuropathology report are also sent to the investigators on request.
Once the diagnosis is complete the specimens are shipped on dry ice (for frozen samples) or in vacuum-sealed plastic (for fixed samples) to the various investigators. All the data regarding the material collected by the brain bank are stored in a database and the confidentiality of the patient data is maintained at all times. The existence of such a database provides instant access to relevant tissue, patient and autopsy data which are registered in a standardized manner. This also facilitates the matching task for both the NBB in supplying the samples and the investigators in setting up their experiments and interpreting the results correctly.
Standardization of protocols for collecting brain bank material:
The samples provided by the bank are well documented and are controlled for quality in order to permit users to draw the accurate meaningful conclusions from their studies. Standardizing the documentation, collection, dissection, processing and storage procedures can ensure these demands. To assure the specimens quality, the bank pays special attention to monitoring the agonal state of the deceased prior to death. Measuring the pH of the brain in CSF collected during rapid autopsies makes it possible to match the samples for agonal state and has therefore been introduced as a routine procedure in the Netherlands Brain Bank. From observations made on human autopsy material collected by the NBB it became evident that the pH value measured in CSF is not affected by the post-mortem delay (Ravid et al., 1992). Tissue pH has been also reported to be a fair indicator of mRNA preservation in human post-mortem brain (Kingsbury et al., 1995). Measuring pH provides a simple means of screening and is thus a crucial measure for agonal state that has to be included in brain banking routine procedures.
Changes in freezing procedures, fixation and storage time may affect many of the parameters used to assess changes in the brain and the potentialities of staining procedures considerably. Tissues used for biochemical studies are usually rapidly frozen and slowly thawed. Synaptosomal preparations from post-mortem tissue examined by electron microscopy showed they were only slightly less pure than preparations from fresh tissue (Hardy et al., 1982). However, to isolate synaptosomes which are morphologically well preserved and have retained their metabolic performance, one should use the opposite procedure as snap-freezing generally yields metabolically and functionally inactive preparations (Hardy et al., 1983).
User survey response and outcome assessment:
Facts and Figures
So far the Netherlands Brain Bank has provided brain material from 2806 autopsies for 600 research projects in countries all over the world (The Netherlands, other European countries, North America, Middle and Far East). Over 600 papers have been published in the international scientific press as an outcome of research using these specimens. The autopsies were performed on patients who suffered from various neurological and psychiatric disorders.
Autopsies 1985 - 2006
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