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The Niedersachsen Live-Tissue and primary cell Bio-Bank (NLTB)


The NLTB project aims at linking state-of-the-art research advances made for the study of host-pathogen and host-allergen interactions, with pre-clinical testing of preventive and therapeutic tools to effectively implement alternative methods to animal experimentation. Many technological advances in fundamental and basic research can contribute significantly to successful alternatives to animal experimentation. However, such technological advances need to be developed in view of standardized and reproducible use, quality assurance and technology transfer. To work toward filling these gaps, NLTB will establish and maintain a repository of protocols, techniques and a biobank of in vitro and ex vivo respiratory cells and tissues for their on-demand delivery and use. The material generated by NLTB will include primary and immortalized cell cultures, air-liquid interface cultures (ALI), tracheal organ cultures (TOC) and precision-cut lung slices (PCLS) derived from a variety of species. The samples will be opportunistically collected at slaughterhouses, from companion animals euthanized for unrelated reasons, surplus experimental animals, and from humans subjected to lung surgery.
Complex interactions between pathogens or allergens and host defence mechanisms are critical determinants of course and severity of infections and allergic reactions, disease outcome and activation of innate and adaptive immune responses. Reproduction of these interactions in laboratory settings as they take place in vivo is essential for the understanding of the pathogenesis of infections and allergies, as well as for pre-clinical screening and eventual testing of the efficacy of therapeutic and preventive drugs. Such studies typically rely on extensive use of laboratory animals, representing the closest models for infections or allergies, mimicking the complexity of the interactions, yet aiming at the generation of reproducible data. The resulting in-depth understanding of these interactions is the basis for the rational development of therapeutic and preventive drugs that target critical steps in the pathogen’s life cycle, allergic sensitisation and detrimental host responses.
As for toxicology studies, the use of in vitro or ex vivo systems, like primary and/or immortalized cell cultures and cell lines, organ cultures and tissue co-cultures, holds promise for the reduction or even replacement of experimental animals in pathogenesis, drug screening, and quality assurance testing. Their multi-replicated use offers the additional advantage of a high statistical power and better reproducibility than the use of experimental animals. The diversity of primary and/or immortalized cell cultures, cell lines and this across organ systems and animal species needs to be extended. Furthermore, existing in vitro and ex vivo systems need to be compared to evaluate the best protocols and approaches to pre-defined research goals. For example, PCLS are a potent ex vivo differentiated epithelial cell model for animal and human lungs: they (i) can be obtained in large numbers, (ii) are largely maintained in their original setting, (iii) are viable for more than a week, (iv) show characteristic functions of the respiratory tract such as mucus-production, ciliary activity, and bronchoconstriction. Application of PCLS has broadened from testing functional responses, like airway and vasoconstrictions, to pharmacological and toxicological testing in various species. However, it has been limitedly applied in basic research, e.g., to elucidate calcium signaling, early allergic responses and viral infection responses.
Other in vitro respiratory systems include ALI cultures and differentiated airway epithelial cells and cell lines. Although these are generally accepted and validated systems to study host-pathogen interactions, their relevance for the replacement of experimental animals needs to be further evaluated.
To this end, the NLTB will have the following objectives:
- Establishment of standardized protocols for the generation of (immortalized) primary respiratory cell cultures, ALI cultures, TOC, and microtome PCLS of slaughtered animals (poultry, swine, cattle), companion animals euthanized for unrelated reasons as well as from surplus experimental animals (poultry, rodents, ferrets, monkeys) and from human surgery
- Establishment of standardized protocols for the cryopreservation of immortalized cell cultures, TOC and PCLS
- Establishment and maintenance of the NLTB
In summary, NLTB aims at establishing and validating protocols and approaches for the replacement and reduction of animal experiments by expanding availability and diversity of live tissues and cell lines, for research on respiratory infections and chronic respiratory diseases of humans and animals. Long-term storage of cells and tissues will facilitate the planning and execution of experiments and allow easy distribution of tissues. NTLB will build on coordinated and efficient logistics for the generation and/or preservation of live tissues and cell cultures, allowing delivery and use of in vitro and ex vivo tissues on demand.



R2N, Niedersächsisches Ministerium für Wissenschaft und Kultur



Prof. Dr. Wolfgang Baumgärtner, PhD (wolfgang.baumgaertner@tiho-hannover.de; 0511-856-8620)

Institut für Pathologie, Stiftung Tierärztliche Hochschule Hannover

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