Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro.
Noroviruses (NoVs) and Sapoviruses (SaVs) are enteric caliciviruses that have been detected in multiple mammalian species, including humans. Currently, efficient cell culture systems have been established only for murine NoVs and porcine SaV Cowden strain. Establishment of an efficient in vitro cell...
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doaj-410c84e7f75e49ee829f005bc670271d2020-11-24T21:38:10ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-01132e017815710.1371/journal.pone.0178157Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro.Tomoichiro OkaGarrett T StoltzfusChelsea ZhuKwonil JungQiuhong WangLinda J SaifNoroviruses (NoVs) and Sapoviruses (SaVs) are enteric caliciviruses that have been detected in multiple mammalian species, including humans. Currently, efficient cell culture systems have been established only for murine NoVs and porcine SaV Cowden strain. Establishment of an efficient in vitro cell culture system for other NoVs and SaVs remains challenging; however, human NoV (HuNoV) replication in 3D cultured Caco-2 cells and a clone of Caco-2 cells, C2BBe1, human enteroids and in human B cells has been reported. In this study, we tested various cells and culture conditions to grow HuNoVs and a human SaV (HuSaV) to test the possibility of the propagation in different cells and culture conditions. We also attempted to grow a bovine NoV (BoNoV) in ex vivo organ cultures. We did not observe significant RNA level increases for HuSaV and BoNoV under our test conditions. HuNoV RNA levels increased to a maximum of ~600-fold in long-term Caco-2 cells that were cultured for 1-2 months in multi-well plates and inoculated with HuNoV-positive and bacteria-free human stool suspensions using serum-free medium supplemented with the bile acid, GCDCA. However, this positive result was inconsistent. Our results demonstrated that HuNoVs, BoNoV and HuSaV largely failed to grow in vitro under our test conditions. Our purpose is to share our findings with other researchers with the goal to develop efficient, reproducible simplified and cost-effective culture systems for human and animal NoVs and SaVs in the future.http://europepmc.org/articles/PMC5810978?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tomoichiro Oka Garrett T Stoltzfus Chelsea Zhu Kwonil Jung Qiuhong Wang Linda J Saif |
spellingShingle |
Tomoichiro Oka Garrett T Stoltzfus Chelsea Zhu Kwonil Jung Qiuhong Wang Linda J Saif Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. PLoS ONE |
author_facet |
Tomoichiro Oka Garrett T Stoltzfus Chelsea Zhu Kwonil Jung Qiuhong Wang Linda J Saif |
author_sort |
Tomoichiro Oka |
title |
Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. |
title_short |
Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. |
title_full |
Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. |
title_fullStr |
Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. |
title_full_unstemmed |
Attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. |
title_sort |
attempts to grow human noroviruses, a sapovirus, and a bovine norovirus in vitro. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2018-01-01 |
description |
Noroviruses (NoVs) and Sapoviruses (SaVs) are enteric caliciviruses that have been detected in multiple mammalian species, including humans. Currently, efficient cell culture systems have been established only for murine NoVs and porcine SaV Cowden strain. Establishment of an efficient in vitro cell culture system for other NoVs and SaVs remains challenging; however, human NoV (HuNoV) replication in 3D cultured Caco-2 cells and a clone of Caco-2 cells, C2BBe1, human enteroids and in human B cells has been reported. In this study, we tested various cells and culture conditions to grow HuNoVs and a human SaV (HuSaV) to test the possibility of the propagation in different cells and culture conditions. We also attempted to grow a bovine NoV (BoNoV) in ex vivo organ cultures. We did not observe significant RNA level increases for HuSaV and BoNoV under our test conditions. HuNoV RNA levels increased to a maximum of ~600-fold in long-term Caco-2 cells that were cultured for 1-2 months in multi-well plates and inoculated with HuNoV-positive and bacteria-free human stool suspensions using serum-free medium supplemented with the bile acid, GCDCA. However, this positive result was inconsistent. Our results demonstrated that HuNoVs, BoNoV and HuSaV largely failed to grow in vitro under our test conditions. Our purpose is to share our findings with other researchers with the goal to develop efficient, reproducible simplified and cost-effective culture systems for human and animal NoVs and SaVs in the future. |
url |
http://europepmc.org/articles/PMC5810978?pdf=render |
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