Characterisation of new animal cell cultures’ sensitivity to Coxsackievirus B5 and Herpes simplex virus‑1

• Published in: Biopreparaty Vol. 23; no. 1; pp. 102 - 110
• Main Authors: Zakharova, Yu. A., Ostapchuk, A. V., Wasielewski, W. W., Fedotova, O. S., Shmeleva, N. A.
• Format: Magazine Article
• Language: English; Russian
• Published: Ministry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products 01.04.2023
• Subjects: coxsackievirus b5; cvb5; diploid animal cell culture; herpes simplex virus-1; hsv-1; porcine foetal kidney fibroblasts; porcine foetal larynx fibroblasts
• ISSN: 2221-996X; 2619-1156
• DOI: 10.30895/2221-996X-2023-23-1-102-110

The increase in the number of cell cultures for virology and biotechnology enhances the chances of a successful response to threats related to outbreaks of well-known and new human infectious diseases. It is a vital task to search for cell cultures sensitive to a wide spectrum of viruses. The aim of the study was to investigate the sensitivity of new diploid animal cell cultures (fibroblasts of a foetal pig’s kidneys and larynx) to Coxsackievirus B5 (CVB5) and Herpes simplex virus-1 (HSV-1). Materials and methods. The cultures of porcine foetal kidney fibroblasts (PFKF) and porcine foetal larynx fibroblasts (PFLF) were derived from a foetus of a healthy pig by mild trypsinisation. The study determined the sensitivity of these new PFKF and PFLF cultures to the above-mentioned viruses by the cytopathic effect (CPE) expressed as a percentage. The infectious activity of CVB5 was studied using real-time polymerase chain reaction (PCR) with the determination of amplification cycle threshold values (C t ); that of HSV-1 was studied using quantitative titration of the virus-containing liquid (VCL). Infectious activity values were expressed as tissue culture 50% infective doses (TCID 50 ). Results. The authors developed diploid PFKF and PFLF cell cultures. PFKF cells demonstrated high sensitivity to CVB5, with a CPE of 87.5±3.3% after passage 3 and a satisfactory concentration of enterovirus RNA in the VCL of 22–24 C t . The sensitivity of PFKF cells to HSV-1 corresponded to a CPE of 92.1±5.5%. In these cells, the infectious activity of HSV-1 corresponded to 10 4.25 TCID 50 /0.2 mL. The experiments with PFLF cells showed low CPE and infectious activity values for both viruses. Conclusions. The study demonstrated high CPE values with the CVB5 (CB5-8100) and HSV-1 (HSV-1/L-2) strains as examples and confirmed the sensitivity of the new diploid PFKF cell culture to these test viruses. Thus, the PFKF cell culture offers potential applications in virology and biotechnology and may be a candidate for testing other strains of CVB5 and HSV-1.