Bulletin "Veterinary biotechnology"

Veterynarna biotehnologija – Veterinary biotechnology, 2018, 32(1), 442-446 [in Ukrainian]. https://doi.org/10.31073/vet_biotech32(1)-59

GAVRASIEVA N.V.e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., KUZMYCH G.S., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

State Scientific Control Institute of Biotechnology and strains (DNKIBSHM)

REQUIREMENTS OF UKRAINE AND EU NORMATIVE DOCUMENTS TO THE WORK OF CONTINUOUS CELL CULTURES IN THE DEPOSIT PROCEDURE

Introduction. The widespread use of cell cultures requires constant monitoring of their biological properties, such as interspecies authenticity, karyotypic stability and purity of contaminating agents.

The goal of the work was to analyze the requirements of SSU, OIE and European Pharmacopoeia to work with cell cultures.

Materials and methods: Normative documents of the EU, OIE and SSU, which regulate requirements for cultures of cells used in veterinary medicine.

Results: It was deposited in 3 cultures of cells (PK-15, Vero, A-72) in the indicators recommended by normative documents at the SSCIBMS. The cultures were grown using DMEM Eagle nutritional media, to which 10% of the embryonic serum of bovine blood (BRL) was added at 370C; pH 7.2–7.4, manufactured by GIBCO and SIGMA. The microscopy was carried out by the formed cell culture monolayer according OIE and the recommendations of the European Pharmacopoeia 5.2.4.. There was no marked contact inhibitors, multicellular cells and any other cellular anomalies, and changes in the color of the nutrient medium; The absence of contamination of bacterial and fungal microflora according to SSU 4483: 2005 and those and recommendations of the European Pharmacopoeia 2.6.1. The investigated samples were not contaminated with bacterial and fungal microflora. The absence of mycoplasma contamination. Monitoring of the absence of contamination with mycoplasmas is carried out according to SSU 4613: 2006 and the recommendations of the European Pharmacopoeia 2.6.7. In the studied samples, the growth of colonies mycoplasma was not observed. In the study of the absence of contamination of foreign viruses in Ukraine, SSU 4517: 2006 is used. Veterinary preparations immuno-biological. Methods of detecting contamination by foreign viruses and recommendations of the European Pharmacopoeia – 5.2.4. During the term of cultivation in the cell monolayer, there were no signs of degeneration caused by an infectious agent. The karyotype of the test is described in the recommendations of the European Pharmacopoeia – section 5.2.4. The karyotypes were identical on 15 passages. Cultures of cells RK-15, Vero, A-72 corresponded to the declared indicators of normative documents SSU and European Pharmacopoeia.

Conclusions and perspectives of further research. According to the recommendations of normative documents of SSU, OIE and European Pharmacopoeia were investigated cell culture. The obtained research results were entered into the culture passport (RK-15, Vero, A-72) and deposited data lines. It is planned to carry out research on other cultures of cells that are in the collection at the SSCIBMS and to conduct a deposit procedure.

Keywords: normative documents, cell culture, deposit.

REFERENCES

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