Disentangling the heterogeneity of multiple sclerosis through identification of independent neuropathological dimensions.

Multiple sclerosis (MS) is a heterogeneous neurological disorder with regards to clinical presentation and pathophysiology. Here, we investigated the heterogeneity of MS by performing an exploratory factor analysis on quantitative and qualitative neuropathology data collected for 226 MS donors in the Netherlands Brain Bank autopsy cohort. Three promising dimensions were identified and subsequently validated with clinical, neuropathological, and genetic data. Dimension 1 ranged from a predominance of remyelinated and inactive lesions to extensive pathological changes, higher proportions of active and mixed lesions, and foamy microglia morphology. This pattern was positively correlated with more severe disease, the presence of B and T cells, and neuroaxonal damage. Scoring high on dimension 2 was associated with active lesions, reactive sites, and the presence of nodules. These donors had less severe disease, a specific pattern of cortical lesions, and MS risk variants in the human leukocyte antigen region, the latter indicating a connection between disease onset and this neuropathological dimension. Donors scoring high on dimension 3 showed increased lesional pathology with relatively more mixed and inactive lesions and ramified microglia morphology. This pattern was associated with longer disease duration, subpial cortical lesions, less involvement of the adaptive immune system, and less axonal damage. Taken together, the three dimensions may represent (1) demyelination and immune cell activity associated with pathological and clinical progression, (2) microglia (re)activity and possibly lesion initiation, and (3) loss of lesion activity and scar formation. Our findings highlight that a thorough understanding of the interplay between multiple pathological characteristics is crucial to understand the heterogeneity of MS pathology, as well as its association with genetic predictors and disease outcomes. The scores of donors on the dimensions can serve as an important starting point for further disentanglement of MS heterogeneity and translation into observations and interventions in living cohorts with MS.

A cell bank paradigm for preclinical evaluation of an analogous cellular product for an allogeneic cell therapy.

Toward the translation of allogeneic cell therapy products, cell banks are needed not only to manufacture the final human product but also during the preclinical evaluation of an animal-based analogous cellular product (ACP). These cell banks need to be established at both the master cell bank (MCB) level and the working cell bank (WCB) level. Inasmuch as most of the development of cell therapy products is at academic centers, it is imperative that academic researchers understand how to establish MCBs and WCBs within an academic environment. To illustrate this process, using articular cartilage as the model, a cell bank for an ACP was developed (MCBs at passage 2, WCBs at passage 5) to produce self-assembled neocartilage for preclinical evaluation (constructs at passage 7). The cell bank system is estimated to be able to produce between 160 000 and 400 000 constructs for each of the six MCBs. Overall, the ACP cell bank yielded constructs that are analogous to the intended human product, which is critical toward conducting preclinical evaluations of the ACP for inclusion in an Investigational New Drug application to the FDA.

Urgent Need for Regulatory Oversight of Human Cells, Tissues, and Cellular and Tissue-Based Products.

This Viewpoint discusses the death of a patient caused by unregulated biological products and efforts to encourage federal government oversight of such products.

Double Chromogen-based Immunohistochemical Staining: An Efficient Approach for Utilizing Long-term Formalin-fixed Tissue in Biobanks.

The New South Wales Brain Tissue Resource Centre is a human brain bank that provides top-quality brain tissue for cutting-edge neuroscience research spanning various conditions from alcohol use disorder to neurodegenerative diseases. However, the conventional practice of preserving brain tissue in formalin poses challenges for immunofluorescent staining primarily due to the formalin's tendency, over time, to create cross-links between antigens, which can obscure epitopes of interest. In addition, researchers can encounter issues such as spectral bleeding, limitations in using multiple colors, autofluorescence, and cross-reactivity when working with long-term formalin-fixed brain tissue. The purpose of the study was to test chromogen-based double immunolabeling to negate the issues with immunofluorescent staining. Colocalization of antigens was explored using chromogens 3-amino-9-ethylcarbazole (AEC) and 3,3,-diaminobenzidine in a sequential staining procedure where the AEC signal was eliminated by alcohol treatment. Combinations of 2 or 3 primary antibodies from the same or different species were trialed successfully with this protocol. The colocalization of antigens was also demonstrated with pseudocoloring that mimicked immunofluorescence staining. This staining technique increases the utility of archival formalin-fixed tissue samples.

Availability of individual proteins for quantitative analysis in postmortem brains preserved in two different brain banks.

AIM: Postmortem brain research is necessary for elucidating the pathology of schizophrenia; an increasing number of studies require a combination of suitable tissue samples preserved at multiple brain banks. In this study, we examined whether a comparative study of protein expression levels can be conducted using postmortem brain samples preserved in different facilities. METHODS: We compared the demographic factors of postmortem brain samples preserved in two institutions and measured and compared the expression levels of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and glial fibrillary acidic protein (GFAP) in the prefrontal cortex and superior temporal gyrus. GAPDH is generally used as a loading control for western blotting, and GFAP is considered as an astrocyte marker in the brain. RESULTS: We found significant differences between the two institutions in postmortem interval, age at death, and preservation time. To reduce the effects of these differences on our measurements, the parameters were set as covariates in our analyses of covariance. Subsequently, no differences in GAPDH and GFAP expression were found between institutions. CONCLUSIONS: When studies are conducted using brain samples preserved in different brain banks, differences in demographic factors should be carefully considered and taken into account by statistical methods to minimize their impact as much as possible. Since there was no significant difference in the protein expression levels of GAPDH and GFAP in either region between the two institutions that preserved the postmortem brains, we concluded that it is possible to perform protein quantitative analysis assuming that there is no effect of difference between two institutions.

Brain banking in the United States and Europe: Importance, challenges, and future trends.

In recent years, brain banks have become valuable resources for examining the molecular underpinnings of various neurological and psychological disorders including Alzheimer disease and Parkinson disease. However, the availability of brain tissue has significantly declined. Proper collection, preparation, and preservation of postmortem autopsy tissue are essential for optimal downstream brain tissue distribution and experimentation. Collaborations between brain banks through larger networks such as NeuroBioBank with centralized sample request mechanisms promote tissue distribution where brain donations are disproportionately lower. Collaborations between brain banking networks also help to standardize the brain donation and sample preparation processes, ensuring proper distribution and experimentation. Ethical brain donation and thorough processing enhances the responsible conduct of scientific studies. Education and outreach programs that foster collaboration between hospitals, nursing homes, neuropathologists, and other research scientists help to alleviate concerns among potential brain donors. Furthermore, ensuring that biorepositories accurately reflect the true demographics of communities will result in research data that reliably represent populations. Implementing these measures will grant scientists improved access to brain tissue, facilitating a deeper understanding of the neurological diseases that impact millions.

Brain Banking in Dementia Studies.

It is now well-established practice in dementia that one clinical entity may be caused by various neurodegenerative disorders, each with different histopathological findings, whereas neuropathologically confirmed patients may have different, unusual, and atypical clinical manifestations.This inconsistency in dementia patients leads to neuropathological examination of cases, and neuropathological examination seems to be an inevitable part of dementia practice, at least until all clinical entities are properly identified for humans.Additionally, the development of disease-modifying therapies and confirmation of the actual accurate diagnosis of the neurodegenerative disease that the drug is thought to modify or act upon are of great importance for neuropathological evaluation in brain banks.Neuropathological processes coexisting among patients diagnosed with established clinical criteria or international guidelines have provided a new perspective in the context of drug development.Here, we review our routinely used methodology in the context of the brain banking process.

Deriving life-course residential histories in brain bank cohorts: A feasibility study.

INTRODUCTION: The exposome is theorized to interact with biological mechanisms to influence risk for Alzheimer's disease but is not well-integrated into existing Alzheimer's Disease Research Center (ADRC) brain bank data collection. METHODS: We apply public data tracing, an iterative, dual abstraction and validation process rooted in rigorous historic archival methods, to develop life-course residential histories for 1254 ADRC decedents. RESULTS: The median percentage of the life course with an address is 78.1% (IQR 24.9); 56.5% of the sample has an address for at least 75% of their life course. Archivists had 89.7% agreement at the address level. This method matched current residential survey methodology 97.4% on average. DISCUSSION: This novel method demonstrates feasibility, reproducibility, and rigor for historic data collection. To our knowledge, this is the first study to show that public data tracing methods for brain bank decedent residential history development can be used to better integrate the social exposome with biobank specimens. HIGHLIGHTS: Public data tracing compares favorably to survey-based residential history. Public data tracing is feasible and reproducible between archivists. Archivists achieved 89.7% agreement at the address level. This method identifies residences for nearly 80% of life-years, on average. This novel method enables brain banks to add social characterizations.

Biological parameters for quality evaluation of allografts from the Brazilian National Institute of Traumatology and Orthopedics tissue bank.

Bone allografts are clinically used in a variety of surgical procedures, and tissue banks are responsible for harvesting, processing, quality testing, storing, and delivering these materials for transplantation. In tissue banks, the bone is processed for the removal of all organic content, remaining only the tissue structure (scaffold). However, several studies have shown that even after using different processing methods, viable cells, functional proteins, and DNA may still persist in the tissue, which constitute the main causes of graft rejection. Therefore, the objective of this study was to establish techniques and biological parameters for quality validation of allografts. To this end, we propose the use of 3 combined methods such as microscopy, histology, and molecular biology techniques to evaluate the quality of allografts harvested and processed by the Brazilian National Institute of Traumatology and Orthopedics (INTO) tissue bank according to the donation criteria of the Brazilian National Health Surveillance Agency and the Brazilian National Transplant System. Bone fragments from different processing stages showed no viable cells on histology, an intact extracellular matrix on scanning electron microscopy, and gradual reduction in DNA amount. Different techniques were used to demonstrate the quality of allografts produced by the INTO tissue bank and to establish biological parameters for ensuring the safety and quality of these products. Future studies need to be undertaken to assess and validate the efficacy of the decellularization process in larger bone grafts with diverse architectural configurations.

Allografts use in orthopedic surgery: trend change over the past 11 years from a regional tissue bank.

Allografts are the second most transplanted tissue in medicine after blood and are now increasingly used for both primary and revision surgery. Allografts have the advantages of lower donor site morbidity, availability of multiple grafts, and shorter operative time. The Banks represents the bridge between Donor and Recipient and guarantees the quality and safety of the distributed allografts Given the increasing interest in these tissues, a retrospective analysis of data collected from the Regional Musculoskeletal Tissue Bank registry over an 11-year period (2009-2019) was conducted. The statistical analyses used were the Shapiro-Wilk normality test and a Poisson regression model. From January 2009 to December 2019, a total of 14,199 musculoskeletal tissues stored in the Regional Musculoskeletal Tissue Bank were provided for surgical allograft procedures. In 2009, the number of allografts performed was 925; this figure has steadily increased to 1599 in 2019. Epiphyses were taken as the reference tissue with an almost constant trend over the period, while a significant increase was denoted for extensor mechanism allograft, ligaments, tendons and long bone corticals (p < 0.001), processed bone tissues had no change in trend (p = 0.841). There was also a gradual decrease in the rate of microbiological positivity, as determined by bacteriological and serological tests performed on the collected tissues. This phenomenon is due to improved sampling techniques and the training of a dedicated team. Thus, we have seen how the use of allografts in orthopedic surgery has increased over the past 11 years, uniformly in terms of tissue type, except for the noticeable increase in ligamentous tissue.

Last twenty-years activity of cardiovascular tissue banking in Barcelona.

The Barcelona Tissue Bank was established from the merge of two previous multi-tissue banks. Potential donors are screened by Donor Center staff and multi-tissue retrieval is performed by specialized own teams. Tissue processing and preservation is performed in clean room facilities by specialised personnel. After quality control of both donor and all tissues results, the heart valves and vascular segments are stored until medical request. The aim of this report is to present the cardiovascular tissue activity and retrospectively evaluate the outcomes of the changes performed in last 20 years. Cardiovascular tissue from 4088 donors was received, specifically 3115 hearts and 2095 vascular segments were processed and evaluated. A total of 48% of the aortic valves, 68% of the pulmonary valves and 75% of the vascular segments were suitable for transplant. The main reason for discarding tissue was macroscopic morphology followed by microbiological results, for both valves and arteries. Altogether, 4360 tissues were distributed for transplantation: 2032 (47%) vascular segments, 1545 (35%) pulmonary valves and 781 (18%) aortic valves. The most common indication for aortic valve surgery was the treatment of endocarditis, while for pulmonary valves, it was congenital malformation reconstruction. Vascular segments were mainly used for reconstruction after ischemia. During this period, a number of changes were made with the goal of enhancing tissue quality, safety and efficacy. These improvements were achieved through the use of a new antibiotic cocktail, increasing of donor age criteria and changing the microbiological control strategy.

The Landscape of US and Global Rare Tumor Research Programs: A Systematic Review.

Rare cancers and other rare nonmalignant tumors comprise 25% of all cancer diagnoses and account for 25% of all cancer deaths. They are difficult to study due to many factors, including infrequent occurrence, lack of a universal infrastructure for data and/or tissue collection, and a paucity of disease models to test potential treatments. For each individual rare cancer, the limited number of diagnosed cases makes it difficult to recruit sufficient patients for clinical studies, and rare cancer research studies are often siloed. As a result, progress has been slow for many of these cancers. While rare cancer research efforts have increased over time, the breadth of the research landscape is not known. A recent literature search revealed a sharp increase in rare tumor, and rare cancer publications began in the early 2000s. To identify rare cancer research efforts being conducted in the US and globally, we conducted an online search of rare tumor/rare cancer research programs and identified 76 programs. To gain a deeper understanding of these programs, we composed and conducted a survey to ask programs for details about their research efforts. Of the 42 programs contacted to complete the survey, 23 programs responded. Survey results show most programs are collecting clinical data, molecular data, and biospecimens, and many are conducting molecular analyses. This landscape analysis demonstrates that multiple rare cancer research efforts are ongoing, and the rare cancer community may benefit from collaboration among stakeholders to accelerate research and improve patient outcomes.

Experience with Tissue Bank Services in 2014 and 2020 in Turku, Finland.

BACKGROUND: The objective of a musculoskeletal tissue bank is to collect, test, store, and provide musculoskeletal tissue allografts required in orthopedic procedures. Strict exclusion criteria are followed when selecting suitable cadaver musculoskeletal tissue donors, and the allografts are procured under sterile conditions to avoid bacterial contamination. Tissue banking in Turku, Finland, began in 1972, and tissue bank services were last reviewed in 2003. This study aimed to review the operation of the musculoskeletal tissue bank in Turku, Finland, between 2014 and 2020 and to analyze the number, types, and contamination rate of the allografts procured from the cadaver donors. Potential donor-related factors causing bacterial contamination of the allografts and whether potential musculoskeletal tissue donors were overlooked among multiorgan donors were also studied. METHODS: A retrospective review of all cadaver musculoskeletal tissue donors used in the Hospital District of Southwest Finland Tyks Orto Musculoskeletal Tissue Bank during the study period was conducted, and data on the procured allograft was collected and presented. The donors were selected among patients treated in the intensive care unit (ICU) of Turku University Hospital (TYKS). RESULTS: A total of 28 cadaver donors were used, and 636 allografts were procured between 2014 and 2020. The bacterial contamination rate was 2.5%, which was lower than that in the previous international literature. The median treatment time in the ICU was significantly longer, and the median value of the highest C-reactive protein level was significantly higher in the group of donors with positive allograft bacterial cultures. CONCLUSIONS: The bacterial contamination rate in the tissue bank was low on an international scale. Some suitable musculoskeletal tissue donors were overlooked among multiorgan donors.

P24-A114†Bringing together the eye banking community throughout Europe and beyond - promoting eye donation in Africa.

PURPOSE: It is estimated that globally there are more than 12.7 million corneal blinds with the vast majority of those living in the developing world. There is huge demand for corneal transplants worldwide as currently only one out of 70 patients can be provided with a cornea.Following the spirit of EEBA in bringing together the international eye banking community we present on our efforts and vision in contributing to the elimination of avoidable blindness in Africa by promoting sustainable eye donation programs. METHODS: At the congress of the South African Tissue Bank Association (SATiBA) in November 2022 a dedicated Round Table Discussion takes place on eye donation in Africa, organized by the World Union of Tissue Banking Associations (WUTBA) together with the Global Alliance of Eye Bank Associations (GAEBA), SATiBA and the German Society for Tissue Transplantation (DGFG). Individuals, national and global players in tissue medicine meet aiming to promote and advocate corneal donation in sub-Saharan Africa to establish patient care that is self-sustaining from within the countries.In preparation for the meeting a questionnaire was completed by the participants to understand the current situation in individual countries: Responses by ophthalmologists, tissue bankers, awareness and tissue donation coordinators from Kenya, Uganda, Nigeria, Ethiopia, and South Africa were evaluated. RESULTS: The survey revealed that all countries are establishing national health acts with references to tissue donation or have them in place with regulations still to be detailed. These are fundamental to strengthen confidence in tissue donation and to start developing donation infrastructures. In all countries there is doubt about donation after death showing the need for advocacy towards the public.The aim of the Round Table is creating a momentum of networking and sharing experience to support the African countries in building local infrastructures and becoming independent from tissue imports in the future. CONCLUSION: What frameworks must exist to successfully establish donation programs in Africa? What help can be provided by countries and organizations that have stable donation programs? These and other questions will be attempted at the Round Table. Bringing together experts, bundling synergies, and creating a momentum to promote cornea donation on social, political, and community level will be a step towards the vision of creating a world in which nobody is needlessly visually impaired.

P10-A128†Trending of contamination rates across NHSBT eye banks.

INTRODUCTION: NHS Blood and Transplant Tissue and Eye Services (TES) is a human multi-tissue, tissue bank supplying tissue for transplant to surgeons throughout the UK. NHSBT has two Eye Banks.NHSBT investigated all our corneas discard due to contamination with the aim to review for any patterns. NHSBT Eye Banks performs initial Microbiology sampling on all Corneas in Corneas in Organ Culture Media at 7 Days. Corneas undergo a 2nd Microbiology sampling the day after the cornea is transferred into dextran median. MATERIALS AND METHODS: Any Microbiology positive media Identified pre-transplant are sent to NHSBT's Microbiology Reference Laboratory (MSL) for Identification. Any organisms which are identified post-dispatch are sent to a Referral Laboratory for rapid Identification and Sensitivity/Specificity Testing. FILTON EYE BANK: Contaminated Corneas in Organ Media: 2018- 28 (0.91%) 2019 -45 (1.10%), 2020- 27 (1.03%), 2021- 39 (1.41%), 2022- 43 (2.1%) (until 15/08/22)Most common Identified Organisms: C.Ablicans C. glabrata C.paraphilotisContaminated In Dextran Pre-Transplant: 2018- 4 (0.17%) 2019 -6 (0.18%), 2020- 9 (0.46%), 2021- 0 (0%), 2022- 3 (0.3%) (until 15/08/22). Most common Identified Organisms: Bacillus speciesContaminated in Dextran Post Transplant: 2018- 0 (0%) 2019 -8 (0.23%), 2020- 2(0.10%), 2021- 2 (0.08%), 2022- 1 (0.11%) (until 15/08/22). Most common Identified Organisms: Bacillus speciesDavid Lucas Eye Bank: Contaminated Corneas in Organ Media: 2020- 20(1.8%), 2021- 37(1.96%), 2022- 21(1.4%) (until 15/08/22). Most common Identified Organisms: C.Ablicans C. glabrata C.KefyrContaminated In Dextran Pre-Transplant: 2020- 6(0.8%), 2021- 2(0.14%), 2022- 1(0.08%) (until 15/08/22). Most common Identified Organisms: Bacillus speciesContaminated in Dextran Post Transplant: 2020- 2 (0.26%), 2021- 1 (0.07%), 2022- 2 (0.16%) (until 15/08/22). Most common Identified Organisms: Bacillus species DISCUSSION: Processes and facilities are of same standard between the two NHSBT Eye Banks and contamination rates are comparable. contamination is only identified in Approx1% of corneas processed. Corneas where growth is identified in Dextran is less than 1% of corneas Issued. Of the positive Microbiology samples identified post-Transplant, were mostly identified as Environmental Bacteria and had no patient impact on patient and assumed to have been contaminated by the operator.

Sourcing and development of tissue for transplantation in reconstructive surgery: A narrative review.

The wealth of allogeneic and xenogeneic tissue products available to plastic and reconstructive surgeons has allowed for the development of novel surgical solutions to challenging clinical problems, often obviating the need to inflict donor site morbidity. Allogeneic tissue used for reconstructive surgery enters the tissue industry through whole body donation or reproductive tissue donation and has been regulated by the FDA as human cells, tissues, and cellular and tissue-based products (HCT/Ps) since 1997. Tissue banks offering allogeneic tissue can also undergo voluntary regulation by the American Association of Tissue Banks (AATB). Tissue prepared for transplantation is sterilized and can be processed into soft tissue or bone allografts for use in surgical reconstruction, whereas non-transplant tissue is prepared for clinical training and drug, medical device, and translational research. Xenogeneic tissue, which is most often derived from porcine or bovine sources, is also commercially available and is subject to strict regulations for animal breeding and screening for infectious diseases. Although xenogeneic products have historically been decellularized for use as non-immunogenic tissue products, recent advances in gene editing have opened the door to xenograft organ transplants into human patients. Herein, we describe an overview of the modern sourcing, regulation, processing, and applications of tissue products relevant to the field of plastic and reconstructive surgery.

Temperature Monitoring of Two Different Thermal Boxes for the Transport of Biological Samples.

BACKGROUND: According to the Resolutions of the Collegiate Board of Directors RDC No. 20/2014, 214/2018, and 707/2022, validation of the temperature of thermal boxes for the transport of biological samples must be based on standardized procedures and tested by the Tissue Banks, guaranteeing safety and quality. Therefore, they can be simulated. Our objective was to monitor and compare the temperature of 2 different coolers while transporting biological samples. METHODS: The following items were packed in each of the 2 different thermal boxes (Box 1: Easy Path; Box 2: Safe Box Polyurethane Vegetal): 6 blood samples (30 mL), one bone tissue sample (200 g), 8 hard ice (Gelox, to keep the temperature <8ºC), and internal and external traceability "Time Stamp" sensors installed for measuring and storing temperature data in real-time. The monitored boxes were placed in the trunk of a bus that traveled an approximate distance of 630 km and were then placed in the trunk of a car, under direct sunlight, until they reached a temperature of 8ºC. RESULTS: In Box 1, the internal temperature was maintained in the range of -7ºC to 8ºC for approximately 26 hours. In Box 2, the internal temperature was maintained in the range of -10ºC to 8ºC for approximately 98 hours and 40 minutes. CONCLUSIONS: We concluded that both coolers, under similar storage conditions, are suitable for transporting biological samples, with Box 2 maintaining the desired temperature for longer.

Characteristics of Emergency Medicine Specimen Bank Participants Compared to the Overall Emergency Department Population.

INTRODUCTION: Biorepositories lack diversity both demographically and with regard to the clinical complaints of patients enrolled. The Emergency Medicine Specimen Bank (EMSB) seeks to enroll a diverse cohort of patients for discovery research in acute care conditions. Our objective in this study was to determine the differences in demographics and clinical complaints between participants in the EMSB and the overall emergency department (ED) population. METHODS: This was a retrospective analysis of participants of the EMSB and the entire UCHealth at University of Colorado Anschutz Medical Center (UCHealth AMC) ED population across three periods: peri-EMSB; post-EMSB; and COVID-19. We compared patients consented to the EMSB to the entire ED population to determine differences in age, gender, ethnicity, race, clinical complaints, and severity of illness. We used chi-square tests to compare categorical variables and the Elixhauser Comorbidity Index to determine differences in the severity of illness between the groups. RESULTS: Between February 5, 2018-January 29, 2022, there were 141,670 consented encounters in the EMSB, representing 40,740 unique patients and over 13,000 blood samples collected. In that same time, the ED saw approximately 188,402 unique patients for 387,590 encounters. The EMSB had significantly higher rates of participation from the following: patients 18-59 years old (80.3% vs 77.7%); White patients (52.3% vs 47.8%), and women (54.8% vs 51.1%) compared to the overall ED population. The EMSB had lower rates of participation from patients ≥70 years, Hispanic patients, Asian patients, and men. The EMSB population had higher mean comorbidity scores. During the six months after Colorado's first COVID-19 case, the rate of consented patients and samples collected increased. The odds of consent during the COVID-19 study period were 1.32 (95% CI 1.26-1.39), and the odds of sample capture were 2.19 (95% CI 2.0-2.41). CONCLUSION: The EMSB is representative of the overall ED population for most demographics and clinical complaints.

Vascular allografts for clinical application in Europe: assessment of 30 years of experience with vascular tissue banking in Brussels.

Vascular tissue banking has been carried out in Brussels for over 30 years in compliance with EU and Swiss tissue banking regulations. A total of 2.765 vascular tissue donations were performed in Belgian, French, Netherlands and Suisse transplant centres: 547(20%), 1.013(37%) and 1.205(43%) during the first, second and third periods, respectively. 85% and 18% increase in donations during the second and third decades compared to previous one, were remarkable. Of the 7.066 evaluated vascular tissues, 2.407(227, 921 and 1.259) were discarded (34.1%), whereas 4.659(523, 1.861 and 2.275) accepted (65.9%) during the respective period. Of the 92 donated veins, 44(47.8%) were discarded and 48(52.2%) accepted. Allografts available for clinical application were stored in vapours of liquid nitrogen. A total of 4.636 allografts were delivered and transplanted for cases of infection (58%), critical limb ischaemia (16%) and congenital cardiac surgery (15%). Thirty veins were implanted. The progressive increases in donations of 20%, 37% and 43% and in transplantations of 20.8%, 34.6% and 45% during the first, second and third periods, respectively, were remarkable. Complications were reported after transplantation and these included acute rejection of two femoral arteries one month after transplantation. We conclude that the donation and transplantation of cryopreserved vascular allografts was stable with a progressive increase over time. Allografts were used predominantly for the treatment of infection, limb salvage for critical ischaemia and for neonates and infants with congenital cardiac malformation. Immune related rejection was observed. This should be a subject of future investigation.