ABSTRACT
Abstract Good cell culture practices are a set of technical and management tools recommended for application in research and service laboratories to guarantee the traceability and reproducibility of in vitro experiments. However, most research laboratories do not have a structured quality assurance system and have difficulties organizing their workflows or even priorities in implementing acceptable laboratory practices. In this study, we applied management and quality assurance tools to define the steps necessary to implement acceptable laboratory practices in the multiuser laboratory of cell culture and establish a cell bank at the Carlos Chagas Institute FIOCRUZ-PR. Our team applied the 5W2H and 5S tools for initial diagnosis and established an action plan to implement and manage the laboratory over two years. Thereby, we defined the scope of laboratory activities, including the demand for establishing a cell bank, the supply of cell lines to internal users, user training, and quality control tests. We also mapped the main activities, establishing their flows and all the necessary documentation to ensure traceability and reproducibility. Additionally, the laboratory was organized in compliance with the 5S principles. In conclusion, using management tools, such as the 5W2H and 5S methods, is a simple and affordable method to organize and manage a cell culture laboratory and can be applied to other research laboratories.
ABSTRACT
Objective:To establish a three-level clinical grade human umbilical cord mesenchymal stem cells (hUC-MSCs) bank, including seed cell bank (SCB), master cell bank (MCB) and working cell bank (WCB), and provide hUC-MSCs with controllable quality for clinical research and application.Methods:247 human umbilical cord tissues were isolated, cultured, amplified, subcultured and frozen in GMP laboratory, and the biological characteristics, safety and stability of hUC-MSCs were tested in accordance with the requirements of relevant quality management control specifications.Results:247 strains of hUC-MSCs were isolated and prepared. The prepared hUC-MSCs have good purity and homogeneity without tumorigenicity, show good differentiation ability in biological efficacy, and have strong immunosuppressive effect in the process of co-culture with immune cells. These cells have passed the quality check of National Institute for Food and Drug Control. In this study, a three-level hUC-MSCs bank was established, and it was included into the National Stem Cell Translational Resource Center.Conclusions:A three-level clinical hUC-MSCs bank was successfully established and preliminarily applied to clinical research, which effectively promoted the standardized development of clinical stem cell resource bank and the clinical transformation and application of stem cells in China.
ABSTRACT
To ensure the consistency of quality in recombinant protein production, the cell bank for biologics should be derived from a single clone. A number of techniques have been used for cloning and assurance from the cellular pool after transfection with a target gene. Here, using CHO cell as an example, we summarize the knowledge and understanding of monoclonality of production cell bank from both industries and regulatory authorities, and propose general considerations on the requirements of monoclonality for clinical trial application and new drug application based on current techniques. Furthermore, we suggest quality control strategies and assessment methods for those cell banks from non-single clones.
ABSTRACT
Objective To explore the possibility of establishing the human adipose-derived mesenchymal stem cells (hADSCs) bank as to provide an alternative source for the seed cells of tissue engineering. Methods The cell surface antigens of the purified, expanded hADSCs and the ones following cryopreservation were detected by flow cytometry, cultured in an induced culture medium to induce the osteogenic and adipogenic differentiation. The specific marker alkaline phosphatase (ALP) in osteogenic special medium was also detected by immunohistochemistry. Results The phenotype and expansion possibility of hADSCs after cryopreservation were remained. It could expand for 10 generations. The doubling time was 48 hours. The 2nd, 6th and 10th generation of hADSCs kept stronger ability of osteogenic and adipogenic differentiation. Conclusion The bank of hADSCs has been incipiently established and can provide eligible seed cells for tissue engineering.
ABSTRACT
Background: Vero cell (ATCC) is from kidney of Blue Monkey in Africa. Because of its strong points such as non tumor form, non exotic virus infection, this cell strain is commonly used for vaccin development in the world. Objective: To determine the quality of vero cell supplying by Japan Polio-myelitis Research Center and WHO vero cell supplying by the Company for Vaccine and Biological Production No.1 in use for develop rota vaccine. Subjects and method:A study was conducted in 2 kinds of vero cell (one ATCC 134 generation supplying by Japan Polio-myelitis Research Center and another WHO ATTC 137 generation supplying by the Company for Vaccine and Biological Production No.1) using standard methods. Results and Conclusion: Both these ATCCs had no exotic agents in generation from 134 to 137. The vero working cell bank for vaccine development has been established by the POLYVAC by using standard methods, in accordance with the WHO regulations. The vero working cells established by POLYVAC had the same quality as that of Vabiotech cell bank. Rota virus strains multiplied well on WHO ATTC 137 generation and ATCC 134 generation supplying by Japan Polio-myelitis Research Center.