Estabelecimento de banco de linhagens celulares aplicadas a pesquisa científica e ensaios toxicológicos in vitro

Abstract

He Carlos Chagas Institute (ICC) is the technical-scientific unit of FIOCRUZ that develops biomedical research in Paraná. The research projects developed at ICC include the evaluation of parasite-host interaction, production of biomolecules, cytotoxicity tests, development of alternative methods to the use of animals, stem cell differentiation, among others. All of these areas have in common the use of cell cultures among their study models. To guarantee the availability of cell cultures, user training, and traceability of biological material for research projects, services, and technological development, the objective of this work was the implementation of an ICC Cell Line Bank following a management and quality control system. For this, 5W2H management tools and 5S System were applied to make the initial diagnosis of the structure and laboratory organization and define the scope of documents and methodologies to be implemented. The definition of cell lines used to establish the cell banks were based on the demand of users. Master and work banks were established for the cell lines: NHDF, V79-4, CHO K1, Balb/c 3T3. All these cells were subjected to quality control tests, including cell viability assays and detection of mycoplasma contamination using the Polymerase Chain Reaction (PCR), DNA staining and a commercial kit. Only cell cultures with cell viability higher than 80% and negative for mycoplasma tests were stored in the master bank and made available to researchers through the work bank. This work enabled the identification of needs related to the consolidation of the laboratory structure and documentation, allowing the elaboration and implementation of the control of safety procedures, training, registration, and reports, among other documents that guarantee the traceability and reliability of the cell culture data. This quality controls strengthens the research projects and improve their reproducibility of scientific results.