Skip to main content
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 1988;11(2):95–100. doi: 10.1007/BF01404139

Cultivation techniques for animal coronaviruses: Emphasis on feline infectious peritonitis virus, canine coronavirus, transmissible gastroenteritis virus, and porcine hemagglutinating encephalomyelitis virus

Roger D Woods 1, Ronald D Wesley 1
PMCID: PMC7088848  PMID: 32214595

Summary

Techniques are described for the growth and characterization of some mammalian coronaviruses. Because of the fastidious nature of their growth requirements, most will replicate only in cells derived from the natural host or a closely related species. Fetal cat cells are used to grow FIPV, and porcine cells are used to grow TGEV and HEV. However, CCV will replicate in both feline and canine cells. Although all four of these viruses prefer to replicate in a cell in the stationary phase of growth, FIPV is able to replicate in an actively growing cell. Each virus causes a cytopathic effect in monolayer cell cultures under agar or media 18 to 72 h postinfection. Primary isolation of each virus from field specimens is difficult, although most can usually be isolated after 1 to 3 blind passages in the cell culture.

Key words: enteric cell lines, isolation, growth requirements, coronavirus

References

  • 1.Adams N. R., Hofstad M. S. Isolation of transmissible enteritis agent of turkeys in avian embryos. Avian Dis. 1971;15:426–433. [PubMed] [Google Scholar]
  • 2.Andries K., Pensaert M. Propagation of hemagglutinating encephalomyelitis virus in porcine cell culture. ZBl. Vet. Med. B. 1980;27:280–290. doi: 10.1111/j.1439-0450.1980.tb01693.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Bass E. P., Sharplee R. L. Coronavirus and gastroenteritis in foals. Lancet. 1975;2(7939):822. doi: 10.1016/S0140-6736(75)80058-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Bhatt P. N., Perey D. H., Jonas A. M. Characterization of the virus of sialodacryoadenitis of rats: a member of the coronavirus group. J. Infect. Dis. 1972;126:123–130. doi: 10.1093/infdis/126.2.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Binn L. N., Marchivicki R. H., Stephanson E. H. Establishment of a canine cell line: derivation, characterization and viral spectrum. Am. J. Vet. Res. 1980;41:855–860. [PubMed] [Google Scholar]
  • 6.Bohl E. H. Vaccination against transmissible gastroenteritis (TGE): Pros and Cons. In: Underdahl N. R., editor. Proc. George A. Young Conf. Lincoln, NE: U. NE Press; 1982. pp. 77–81. [Google Scholar]
  • 7.Boursnell M. E. G., Brown T. D. K., Foulds I. J., et al. The complete nucleotide sequence of avian infectious bronchitis virus: analysis of the polymerase-coding region. Adv. Exp. Med. Biol. 1987;218:15–30. doi: 10.1007/978-1-4684-1280-2_3. [DOI] [PubMed] [Google Scholar]
  • 8.Cavanagh D. Structural polypeptides of coronavirus IBV. J. Gen. Virol. 1981;53:93–103. doi: 10.1099/0022-1317-53-1-93. [DOI] [PubMed] [Google Scholar]
  • 9.Crandell R. A., Fabricant C. G., Nelson-Rees W. A. Development, characterization and viral susceptibility of a feline (Felis catus) renal cell line (CrFK) In Vitro. 1973;9:176–185. doi: 10.1007/BF02618435. [DOI] [PubMed] [Google Scholar]
  • 10.Davis H., Macnaughton M. R. Comparison of the morphology of three coronaviruses. Arch. Virol. 1979;59:25–33. doi: 10.1007/BF01317891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Edwards B. G., Fulker R. H., Acree W. M., et al. Evaluating a canine coronavirus vaccine through antigen extinction and challenge studies. Vet. Med. 1985;80:28–33. [Google Scholar]
  • 12.Frederick G. T., Bohl E. H. Local and systemic cell mediated immunity against transmissible gastroenteritis; an intestinal infection of swine. J. Immunol. 1976;116:1000–1004. [PubMed] [Google Scholar]
  • 13.Hiria K., Hitchner S. B., Calnek B. W. Characterization of a new coronavirus-like agent isolated from parrots. Avian Dis. 1979;23:515–525. [PubMed] [Google Scholar]
  • 14.Holmes K. V. Replication of coronaviruses. In: Field B. N., editor. Virology. New York, NY: Raven Press; 1985. pp. 1331–1342. [Google Scholar]
  • 15.Jacobs L., deGroot R., van der Zeijst B. A. M., et al. The nucleotide sequence of the peplomer gene of porcine transmissible gastroenteritis virus (TGEV): comparison with the sequence of the peplomer protein of feline infectious peritonitis virus (FIPV) Virus Res. 1987;8:363–371. doi: 10.1016/0168-1702(87)90008-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Jimeniz G., Correa I., Melgosa M., et al. Critical epitopes in transmissible gastroenteritis virus neutralization. J. Virol. 1986;60:131–139. doi: 10.1128/jvi.60.1.131-139.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Kapke P. A., Brian D. A. Sequence analysis of the porcine transmissible gastroenteritis coronavirus nucleocapsid protein. Virology. 1986;151:41–49. doi: 10.1016/0042-6822(86)90102-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Kapke P. A., Tung F. Y. C., Brian D. A., et al. Nucleotide sequence of the porcine transmissible gastroenteritis coronavirus matrix protein gene. Adv. Exp. Med. Biol. 1987;218:117–122. doi: 10.1007/978-1-4684-1280-2_13. [DOI] [PubMed] [Google Scholar]
  • 19.Kemeny L. J. Isolation of transmissible gastroenteritis virus from pharyngeal swabs obtained from sows at slaughter. Am. J. Vet. Res. 1978;39:703–705. [PubMed] [Google Scholar]
  • 20.Lai M. M. C. Overview: molecular biology of coronaviruses 1986. Adv. Exp. Med. Biol. 1987;218:7–14. [Google Scholar]
  • 21.Macnaughton M. R., Madge M. H. The genome of human coronavirus strain 229E. J. Gen. Virol. 1978;39:497–504. doi: 10.1099/0022-1317-39-3-497. [DOI] [PubMed] [Google Scholar]
  • 22.McClurkin A. W., Norman J. O. Studies on transmissible gastroenteritis of swine. II. Selected characteristics of a cytopathogenic virus common to five isolates from transmissible gastroenteritis. Can. J. Comp. Vet. Sci. 1966;30:190–198. [PMC free article] [PubMed] [Google Scholar]
  • 23.Mebus C. A., Stair E. L., Rhodes M. B., et al. Neonatal calf diarrhoea; propagation, attenuation, and characterization of a coronavirus-like agent. Am. J. Vet. Res. 1973;34:145–150. [PubMed] [Google Scholar]
  • 24.Mengeling W. L. Hemadsorption plaque assay for hemagglutinating encephalomyelitis virus. Am. J. Vet. Res. 1972;33:2075–2080. [PubMed] [Google Scholar]
  • 25.Mengeling W. L., Boothe G. D., Ritchie A. E. Characteristics of a coronavirus (strain 67N) of pigs. Am. J. Vet. Res. 1972;33:297–308. [PubMed] [Google Scholar]
  • 26.Moxley R. A., Olsen L. D. Comparison of TGEV vaccines. Proc. Int. Vet. Soc. Cong. Barcelona, Spain: Int. Pig Vet. Soc. 1986;9:191. [Google Scholar]
  • 27.Payne C. M., Ray C. G., Borduin V., et al. An eight-year study of the viral agents of acute gastroenteritis in humans: ultrastructural observations and seasonal distribution with a major emphasis on coronavirus. Adv. Exp. Med. Biol. 1987;218:579–580. doi: 10.1007/978-1-4684-1280-2_74. [DOI] [PubMed] [Google Scholar]
  • 28.Pedersen N. C., Black J. W. Attempted immunization of cats against feline infectious peritonitis; using avirulent live virus or sublethal amounts of virulent virus. Am. J. Vet. Res. 1983;44:229–234. [PubMed] [Google Scholar]
  • 29.Pedersen N. C., Boyle J. F., Floyd K. Infection studies in kittens using feline infectious peritonitis virus propagated in cell culture. Am. J. Vet. Res. 1981;42:363–367. [PubMed] [Google Scholar]
  • 30.Pedersen N. C., Ward J., Mengeling W. L. Antigenic relationship of the feline infectious peritonitis virus to coronaviruses of other species. Arch. Virol. 1978;58:45–53. doi: 10.1007/BF01315534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Pensaert M., Callebaut P., Vergote J. Isolation of a porcine respiratory, non-enteric coronavirus related to transmissible gastroenteritis. Vet. Qt. 1986;8:257–261. doi: 10.1080/01652176.1986.9694050. [DOI] [PubMed] [Google Scholar]
  • 32.Rasschaert D., Laudv H. The predicted primary structure of the peplomer protein E2 of the porcine coronavirus transmissible gastroenteritis virus. J. Gen. Virol. 1987;68:1883–1890. doi: 10.1099/0022-1317-68-7-1883. [DOI] [PubMed] [Google Scholar]
  • 33.Robb J. A., Bond C. W. Coronaviridae. In: Fraenkel-Conrat H., Wagner R. R., editors. Comprehensive Virology, vol. 14. New York, NY: Plenum Press; 1979. pp. 193–247. [Google Scholar]
  • 34.Schmidt O. W., Cooney M. K., Kenny G. E. Plaque assay and improved yield of human coronavirus in a human rhabdosarcoma cell line. J. Clin. Microbiol. 1979;9:722–728. doi: 10.1128/jcm.9.6.722-728.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Siddell S., Wege H., ter Meulen V. The structure and replication of coronaviruses. Curr. Top. Microbiol. Immunol. 1982;99:131–163. doi: 10.1007/978-3-642-68528-6_4. [DOI] [PubMed] [Google Scholar]
  • 36.Siddell S., Wege H., ter Meulen V. The biology of coronaviruses. J. Gen. Virol. 1983;64:761–776. doi: 10.1099/0022-1317-64-4-761. [DOI] [PubMed] [Google Scholar]
  • 37.Small J. D., Woods R. D. Relatedness of rabbit coronavirus to other coronaviruses. Adv. Exp. Med. Biol. 1987;218:521–528. doi: 10.1007/978-1-4684-1280-2_68. [DOI] [PubMed] [Google Scholar]
  • 38.Stark S. L., Fernelius A. L., Boothe G. D., et al. Transmissible gastroenteritis (TGE) of swine: effect of age of swine testes cell culture monolayers on plaque assays of TGE virus. Can. J. Comp. Med. 1975;39:466–468. [PMC free article] [PubMed] [Google Scholar]
  • 39.Storz J., Rott R., Kaluza G. Enhancement of plaque formation and cell fusion of an enteric pathogenic coronavirus by trypsin treatment. Infect. Immunol. 1981;31:1212–1222. doi: 10.1128/iai.31.3.1214-1222.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Sturman L. S., Holmes K. V. The molecular biology of coronaviruses. Adv. Virus. Res. 1983;28:35–112. doi: 10.1016/S0065-3527(08)60721-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Wege H., Siddell S., ter Meulen V. The biology and pathogenesis of coronaviruses. Curr. Top. Microbiol. Immunol. 1982;99:165–200. doi: 10.1007/978-3-642-68528-6_5. [DOI] [PubMed] [Google Scholar]
  • 42.Woods R. D. Small plaque variant transmissible gastroenteritis virus. J. Am. Vet. Med. Assoc. 1978;173:643–647. [PubMed] [Google Scholar]
  • 43.Woods R. D. Studies of enteric coronaviruses in a feline cell line. Vet. Microbiol. 1982;7:427–435. doi: 10.1016/0378-1135(82)90059-1. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Tissue Culture Methods are provided here courtesy of Nature Publishing Group

RESOURCES