Application of molecular markers for monitoring the quality of human and non-human cell lineages

• Grant number: 22/07783-0
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: February 01, 2023
• End date: October 31, 2023
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Érika Machado de Salles
• Grantee: Érika Machado de Salles
• Company: BS Análises Laboratoriais Ltda
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais ; Atividades profissionais, científicas e técnicas não especificadas anteriormente
• City: São Paulo
• Pesquisadores principais: Eduardo Leal Rodrigues
• Associated researchers: Paula Carolina de Souza
• Associated scholarship(s): 23/01259-0 - Validation of Fluorescent Multiplex PCR amplification assays for non-human cell lineage authentication and mycoplasma detection, BP.TT; 23/01255-4 - Application of molecular markers to monitor the quality of human and non-human cell lineages, BP.PIPE

Abstract

Primary cell cultures, continuous (immortalized) cell lines and tissues are extremely important for the pharmaceutical industry and biomedical research, as they are models for studying more complex physiological systems. Quality control of cell lines is essential to ensure reproducibility. Although cell lines have been used for decades, misidentifications such as line switches and cross-contamination (intra- and interspecific) remain a problem. Thus, tools for authenticating cell lines are critical for quality control in cell-based biological experiments. Current methods to authenticate human cell lines use short tandem repeat markers (STRs), based on procedures successfully used in the forensic community for human identification. However, there are no STR-based methods to authenticate non-human cell lines to date. Therefore, our project aims to validate STR-type genetic markers for authentication of murine cell lines and cells isolated from the kidney of the African green monkey (Chlorocebus aethiop sabaeus), a lineage designated as Vero and widely disseminated in research that cultivates viruses, such as for the isolation of SARS-CoV-2, vaccine development and cytotoxicity assays. In addition, our goal is to develop a cell line authentication service in conjunction with the analysis of constantly tested culture contaminants, such as the presence of mycoplasma and/or the presence of murine genetic material in human cell lines. In an innovative way in Brazil, our entire service will be validated using samples on filter paper and transport at room temperature. In this way, researchers from anywhere in Brazil will have the opportunity to follow up their strains for quality control of their experiments or to confirm the identity of strains received in their laboratory. Our research group seeks to emphasize, in educational terms, the importance of monitoring cell lines, since such tools consist of units with population dynamism, that is, with the advancement of passages, it tends to acquire changes (mutations) that can be neutral. Or that may impact the uniformity of the system, consequently, generating impacts on the reproducibility of the data, which indirectly generates more financial costs and efforts with repetitions. (AU)