Abstract
BackgroundBats 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.Methodology/FindingsBlack 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/SignificanceThe 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.
Highlights
There is increasing evidence to indicate that bats play a major role in the emergence and transmission of new and deadly zoonotic viruses [1]
The simplicity of Method 2 and its reproducibility led to the adoption of this method for our primary cell culture production
With the increasing trend of bat borne viruses crossing the species barrier and causing severe disease in humans and other animals, there is an urgent need for the establishment of cell lines from various bat species to facilitate virus isolation and basic research into virus-host interaction
Summary
There is increasing evidence to indicate that bats play a major role in the emergence and transmission of new and deadly zoonotic viruses [1]. Bats are the putative natural reservoir hosts for a number of emerging zoonotic viruses including Nipah virus (NiV) [2], Hendra virus (HeV) [3], severe acute respiratory syndrome coronavirus (SARS-CoV) [4,5] and Ebola virus [6]. These agents are among some of the most virulent pathogens to emerge from animal reservoirs and are capable of infecting a broad range of species. 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
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