Journal Article

Establishment, Immortalisation and Characterisation of Pteropid Bat Cell Lines

Middleton, Deborah, Michalski, Wojtek P., Virtue, Elena R., Marsh, Glenn A., McEachern, Jennifer A., Grimley, Samantha, Wang, Lin-Fa, Tachedjian, Mary, De Jong, Carol, Liu, Jun-Ping, Field, Hume E., Crameri, Gary, Todd, Shawn, Bulach, Dieter, Smith, Craig, Yu, Meng

2009

Overview

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. 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. 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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Animals / Antigen T (large) / Antigens / Bats / Bats (Animals) / Biotechnology / Cell Biology / Cell culture / Cell Culture Techniques - methods / Cell cycle / Cell division / Cell Line, Transformed - cytology / Cell lines / Cell Shape - drug effects / Cells (Biology) / Chiroptera / Cloning, Molecular / Coronaviridae / Coronaviruses / Culture media / Ebola virus / Fibroblasts / Fisheries / Foxes / Growth factors / Hendra virus / Hendra Virus - drug effects / Hendra Virus - physiology / Henipavirus / Henipavirus Infections - virology / Humans / Immunity, Innate - drug effects / Immunity, Innate - immunology / Immunology / Immunology/Innate Immunity / Infections / Infectious Diseases / Interferons - genetics / Laboratories / Ligands / Livestock / Media (culture) / Monkeys / Nipah virus / Nipah Virus - drug effects / Nipah Virus - physiology / Organs / Pathogens / Poly I-C - pharmacology / Pteropus alecto / RNA-directed DNA polymerase / SARS coronavirus / Severe acute respiratory syndrome / Simian virus 40 / Simian virus 40 - genetics / Stem cells / Telomerase / Telomerase reverse transcriptase / Tumor antigens / Virology / Viruses / Zoonoses