Establishment, immortalisation and characterisation of pteropid bat cell lines.
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 isola...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2009-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2788226?pdf=render |
| _version_ | 1811270570691526656 |
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| author | Gary Crameri Shawn Todd Samantha Grimley Jennifer A McEachern Glenn A Marsh Craig Smith Mary Tachedjian Carol De Jong Elena R Virtue Meng Yu Dieter Bulach Jun-Ping Liu Wojtek P Michalski Deborah Middleton Hume E Field Lin-Fa Wang |
| author_facet | Gary Crameri Shawn Todd Samantha Grimley Jennifer A McEachern Glenn A Marsh Craig Smith Mary Tachedjian Carol De Jong Elena R Virtue Meng Yu Dieter Bulach Jun-Ping Liu Wojtek P Michalski Deborah Middleton Hume E Field Lin-Fa Wang |
| author_sort | Gary Crameri |
| collection | DOAJ |
| description | 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. METHODOLOGY/FINDINGS: 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/SIGNIFICANCE: 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. |
| first_indexed | 2024-04-12T22:04:13Z |
| format | Article |
| id | doaj.art-b99bbc09d8614821bbe48f1105cac65e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T22:04:13Z |
| publishDate | 2009-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-b99bbc09d8614821bbe48f1105cac65e2022-12-22T03:15:00ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-01-01412e826610.1371/journal.pone.0008266Establishment, immortalisation and characterisation of pteropid bat cell lines.Gary CrameriShawn ToddSamantha GrimleyJennifer A McEachernGlenn A MarshCraig SmithMary TachedjianCarol De JongElena R VirtueMeng YuDieter BulachJun-Ping LiuWojtek P MichalskiDeborah MiddletonHume E FieldLin-Fa WangBACKGROUND: 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. METHODOLOGY/FINDINGS: 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/SIGNIFICANCE: 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.http://europepmc.org/articles/PMC2788226?pdf=render |
| spellingShingle | Gary Crameri Shawn Todd Samantha Grimley Jennifer A McEachern Glenn A Marsh Craig Smith Mary Tachedjian Carol De Jong Elena R Virtue Meng Yu Dieter Bulach Jun-Ping Liu Wojtek P Michalski Deborah Middleton Hume E Field Lin-Fa Wang Establishment, immortalisation and characterisation of pteropid bat cell lines. PLoS ONE |
| title | Establishment, immortalisation and characterisation of pteropid bat cell lines. |
| title_full | Establishment, immortalisation and characterisation of pteropid bat cell lines. |
| title_fullStr | Establishment, immortalisation and characterisation of pteropid bat cell lines. |
| title_full_unstemmed | Establishment, immortalisation and characterisation of pteropid bat cell lines. |
| title_short | Establishment, immortalisation and characterisation of pteropid bat cell lines. |
| title_sort | establishment immortalisation and characterisation of pteropid bat cell lines |
| url | http://europepmc.org/articles/PMC2788226?pdf=render |
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