title: Establishment, immortalisation and characterisation of pteropid bat cell lines.
abstract: BACKGROUND
  Bats are the suspected natural reservoir hosts for a number of new and emerging zoonotic viruses including Nipah virus, Hendra virus, severe acute respiratory syndrome coronavirus and Ebola virus. Since the discovery of SARS-like coronaviruses in Chinese horseshoe bats, attempts to isolate a SL-CoV from bats have failed and attempts to isolate other bat-borne viruses in various mammalian cell lines have been similarly unsuccessful. New stable bat cell lines are needed to help with these investigations and as tools to assist in the study of bat immunology and virus-host interactions.
  
  
  RESULTS
  Black flying foxes (Pteropus alecto) were captured from the wild and transported live to the laboratory for primary cell culture preparation using a variety of different methods and culture media. Primary cells were successfully cultured from 20 different organs. Cell immortalisation can occur spontaneously, however we used a retroviral system to immortalise cells via the transfer and stable production of the Simian virus 40 Large T antigen and the human telomerase reverse transcriptase protein. Initial infection experiments with both cloned and uncloned cell lines using Hendra and Nipah viruses demonstrated varying degrees of infection efficiency between the different cell lines, although it was possible to infect cells in all tissue types.
  
  
  CONCLUSIONS
  The approaches developed and optimised in this study should be applicable to bats of other species. We are in the process of generating further cell lines from a number of different bat species using the methodology established in this study.
authors.affiliation: CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Australia.
authors.forename: Gary
authors.initials: G
authors.lastname: Crameri
authors.forename: Shawn
authors.initials: S
authors.lastname: Todd
authors.forename: Samantha
authors.initials: S
authors.lastname: Grimley
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: McEachern
authors.forename: Glenn A
authors.initials: GA
authors.lastname: Marsh
authors.forename: Craig
authors.initials: C
authors.lastname: Smith
authors.forename: Mary
authors.initials: M
authors.lastname: Tachedjian
authors.forename: Carol
authors.initials: C
authors.lastname: De Jong
authors.forename: Elena R
authors.initials: ER
authors.lastname: Virtue
authors.forename: Meng
authors.initials: M
authors.lastname: Yu
authors.forename: Dieter
authors.initials: D
authors.lastname: Bulach
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Liu
authors.forename: Wojtek P
authors.initials: WP
authors.lastname: Michalski
authors.forename: Deborah
authors.initials: D
authors.lastname: Middleton
authors.forename: Hume E
authors.initials: HE
authors.lastname: Field
authors.forename: Lin-Fa
authors.initials: LF
authors.lastname: Wang
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002461:Cell Line, Transformed / Q000166:cytology*; D048430:Cell Shape / Q000187:drug effects; D002685:Chiroptera; D003001:Cloning, Molecular; D045404:Hendra Virus / Q000187:drug effects / Q000502:physiology; D045464:Henipavirus Infections / Q000821:virology; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects / Q000276:immunology; D007372:Interferons / Q000235:genetics; D045405:Nipah Virus / Q000187:drug effects / Q000502:physiology; D011070:Poly I-C / Q000494:pharmacology; D013539:Simian virus 40 / Q000235:genetics
chemical_list: D007372:Interferons; D011070:Poly I-C