Preview

Veterinary Science Today

Advanced search

Studies of biological properties of continuous suspension ВНК-21/SUSP/ARRIAH cell line

https://doi.org/10.29326/2304-196X-2021-3-38-230-238

Abstract

The results of the studies of cytomorphological, karyological, cultural properties of continuous suspension ВНК-21/SUSP/ARRIAH subline of newborn Syrian hamster kidney cells intended for foot-and-mouth disease, rabies, bovine parainfluenza-3, Aujeszky’s disease virus reproduction, as well as for production of diagnostic veterinary biologicals are presented. When cultured in suspension, BHK-21/SUSP/ARRIAH cell subline undergoes selection towards hypoploidy: modal class is represented by cells with 41 chromosomes (32–40% of cells); the share of cells containing 40–42 chromosomes is 78–80%; the share of polyploids averages around 1%; the range of variation in the number of chromosomes is from 36 to 54. BHK-21/SUSP/ARRIAH cell subline cultured in suspension with cell seeding concentration of 0.6–0.8 million cells/cm3 demonstrates growth rate of 6.67–11.00 and 96–99% cell viability. After 48 hours, G1-phase (diploid-2n) cells prevail in the cell population of the new subline (30.0–75.0% of cells); cells that undergo preparation for mitosis (S-phase) and mitosis (G2+M-phase) account for 3.0 to 20.0% of the entire population; the number of meganucleated and multinucleated cells (>4n) at the beginning and at the end of the logarithmic phase increases to 2%. BHK-21/SUSP/ARRIAH cells recover rapidly after cryopreservation and demonstrate 95–99% viability and growth rate of 3.36–5.88 at passages 1 to 3 and 6.85–10.95 at passages 4 to 12. Continuous suspension BHK-21/SUSP/ARRIAH cell line ensures virus accumulation at the following titres: FMD virus – 7.30– 8.00 lg TCID50/cm3, rabies virus – 7.25–8.00 lg CCID50/cm3, bovine parainflunza-3 virus – at least 6.00 lg TCID50/cm3, Aujeszky’s disease virus – 7.50–7.80 lg TCID50/cm3.

About the Authors

M. I. Doronin
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Maksim I. Doronin, Candidate of Science (Biology), Leading Researcher, Laboratory for FMD Prevention

600901, Vladimir, Yur’evets



M. N. Guseva
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Marina  N.  Guseva, Candidate of Science (Biology), Senior Researcher, Laboratory for FMD Prevention

Vladimir



D. V. Mikhalishin
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Dmitry V. Mikhalishin, Candidate of Science (Veterinary Medicine), Head of Laboratory for FMD Prevention

Vladimir



A. S. Sharypov
FSBI “Central Scientific and Methodological Veterinary Laboratory” (FSBI CNMVL)
Russian Federation

Andrey S. Sharypov, Candidate of Science (Veterinary Medicine), Researcher, Department for Research Coordination

Moscow



N. S. Mudrak
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Natalia S. Mudrak, Doctor of Science (Biology), Chief Researcher, Reference Laboratory for Avian Viral Diseases

Vladimir



N. Ye. Kamalova
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Natalia Ye. Kamalova, Doctor of Science (Veterinary Medicine), Chief Researcher, Centre for Preclinical Studies

Vladimir



B. L. Manin
FGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)
Russian Federation

Boris L. Manin, Candidate of Science (Biology), Leading Researcher, Sector for Cell Culture, Innovation Department

Vladimir



References

1. Stoker M., Macpherson I. Syrian hamster fibroblast cell line BHK-21 and its Derivatives. Nature. 1964; 203: 1355–1357. DOI: 10.1038/2031355A0.

2. Manin B. L., Mikhalishin V. V., Korpusova T. I., Stegniy B. T., Lavrik A. A. Biological tolerance parameters of production ВНК-21 cell culture [Parametry biologicheskoj tolerantnosti proizvodstvennoj kletochnoj kul’tury BHK-21]. Veterinary Medicine: Inter-Departmental Subject Scientific Collection. Kharkiv; 2002; 80: 404–408. (in Russian).

3. Sokova V. V., Aliyeva L. А., Bondarenko A. F., Khudyakov G. A. ВНК-21/ 2-17b line of newborn Syrian hamster kidney cells [Kletochnaya liniya pochki novorozhdennogo sirijskogo homyachka BHK-21/2-17b]. Certificate of Authorship No. 240289 USSR. All-Union Foot-and-Mouth Disease Research Institute; 1986. (in Russian).

4. Eliseev A. K., Melnik N. V., Zenov N. I., Litenkova I. Yu., Bogach V. N., Ivanov V. S., et al. BHK-21/13-02-transplantable monolayer-suspension subline of newborn syrian hamster kidney cells, intended for FMD virus and rabies virus reproduction. Patent No. 2614074 Russian Federation, Int. C12N 5/07 (2010.01). FKP “Shchelkovskij biokombinat”. No. 2005116301. Date of filing: 31.05.2005. Date of publication: 22.03.2017. Bull. No. 9. (in Russian).

5. Eliseev A. K., Zenov N. I., Krasutkin S. N., Mel’nik N. V., Khajkina L. S., Litenkova I. Yu., et al. BHK-21/13-13-finite monolayer-suspension cell subline of kidney of new-born syrian hamster, intended for reproduction of foot-and-mouth disease virus and rabies virus. Patent No. 2553552 Russian Federation, Int. C12N 5/071 (2010.01). FKP “Shchelkovskij biokombinat”. No. 2014131866/10. Date of filing: 01.08.2014. Date of publication: 20.06.2015. Bull. No. 17. (in Russian).

6. Russian Cell Culture Collection (RCCC) [Rossijskaya kollekciya kletochnyh kul’tur (RKKK)]: catalogue. Ed. by G. P. Pinayev, G. G. Polyanskaya. St. Petersburg; 2004. 315 p. (in Russian).

7. Bailey J. E., Ollis D. F. Biochemical Engineering Fundamentals. 2nd ed. New Delhi: McGraw-Hill, Inc.; 1986. 984 p.

8. Cell culture techniques [Metody kul’tivirovaniya kletok]: Proceedings of the Academy of Sciences of the USSR. Executive ed. G. P. Pinayev. L.: Nauka; 1988. 319 p. (in Russian).

9. Catalogue of Russian Cell Culture Collection [Katalog Rossijskoj kollekcii kletochnyh kul’tur]. Executive ed. G. P. Pinayev, G. G. Polyanskaya. Omsk: Omsk State Pedagogical University (OSPU); 1999. 429 p. (in Russian).

10. Yurkov S. G., Zuyev V. V., Sidorov S. I., Kushnir S. D., Smyslova N. Yu., Neverovskaya N. S., et al. Catalogue of Cell Culture Collection of the All-Russian Research Institute of Veterinary Virology and Microbiology (VNIIVViM) [Katalog kollekcii kletochnyh kul’tur VNIIVViM]. Pokrov; 2000; 60–62. eLIBRARY ID: 21632428. (in Russian).

11. Lozovoy D. A., Guseva M. N., Mikhalishin D. V., Doronin M. I., Manin B. L., Shishkova A. A., et al. BHK-21/SUSP/ARRIAH – continuous suspension subline of newborn syrian hamster kidney cells, intended for reproduction of foot-and-mouth disease viruses, rabies, parainfluenza- 3, Aujeszky’s disease in producing antiviral vaccines, as well as for making diagnostic and preventive veterinary biopreparations. Patent No. 2722671 Russian Federation, Int. C12N 5/10 (2006.01). FGBI “ARRIAH”. No. 2019131190. Date of filing: 01.10.2019. Date of publication: 02.06.2020. Bull. No. 16. (in Russian).

12. Polyanskaya G. G. Karyotype variation patterns in cell cultures during long-term cultivation in different cell lines [Zakonomernosti kariotipicheskoj izmenchivosti v kletochnyh kul’turah pri dlitel’nom kul’tivirovanii v raznyh liniyah]: author’s abstract of Candidate of Science (Biology) thesis. St. Petersburg; 2000. 38 p. (in Russian).

13. Moorhead P. S., Nowell P. C., Mellman W. J., Battips D. M., Hungerford D. A.

14. Chromosome preparations of leukocytes cultured from human peripheral

15. blood. Exp. Cell. Res. 1960; 20: 613–616. DOI: 10.1016/0014-4827(60)90138-5.

16. Mamayeva S. Ye. Chromosomal analysis of cultured cells [Hromosomnyj analiz kul’tiviruemyh kletok]. In: Cell culture techniques. L.: Nauka; 1987; 78–98. (in Russian).

17. Kudryavtsev I. V., Khaydukov S. V., Zurochka A. V., Chereshnev V. A. Flow cytometry in experimental biology [Protochnaya citometriya v eksperimental’noj biologii]. Yekaterinburg: Ural Branch of the Russian Academy of Sciences (UB RAS); 2012. 192 p. (in Russian).

18. Ormerod M. G., Tribukait B., Giaretti W. Consensus report of the task force on standardisation of DNA flow cytometry in clinical pathology. Analytical Cellular Pathology. 1998; 17 (2): 103–110. DOI: 10.1155/1998/842306.

19. Rais Y., Zviran A., Geula S., Gafni O., Chomsky E., Viukov S., et al. Deterministic direct reprogramming of somatic cells to pluripotency. Nature. 2013; 502 (7549): 65–70. DOI: 10.1038/nature12587.


Review

For citations:


Doronin M.I., Guseva M.N., Mikhalishin D.V., Sharypov A.S., Mudrak N.S., Kamalova N.Ye., Manin B.L. Studies of biological properties of continuous suspension ВНК-21/SUSP/ARRIAH cell line. Veterinary Science Today. 2021;10(3):230-238. https://doi.org/10.29326/2304-196X-2021-3-38-230-238

Views: 527


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 2304-196X (Print)
ISSN 2658-6959 (Online)