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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Health Sciences / Faculty of Health Sciences >
Peer-reviewed Journal Articles, etc >
Subtle changes in host cell density cause a serious error in monitoring of the intracellular growth of Chlamydia trachomatis in a low-oxygen environment: Proposal for a standardized culture method
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| Title: | Subtle changes in host cell density cause a serious error in monitoring of the intracellular growth of Chlamydia trachomatis in a low-oxygen environment: Proposal for a standardized culture method |
| Authors: | Sakai, Kouhei Browse this author | | Matsuo, Junji Browse this author →KAKEN DB | | Watanabe, Takanori Browse this author | | Okubo, Torahiko Browse this author | | Nakamura, Shinji Browse this author →KAKEN DB | | Yamaguchi, Hiroyuki Browse this author →KAKEN DB |
| Keywords: | Chlamydia trachomatis | | Low-oxygen environment | | Host cell density | | RPMI | | DMEM | | CTGF |
| Issue Date: | Oct-2018 |
| Publisher: | Elsevier |
| Journal Title: | Journal of microbiological methods |
| Volume: | 153 |
| Start Page: | 84 |
| End Page: | 91 |
| Publisher DOI: | 10.1016/j.mimet.2018.09.007 |
| PMID: | 30240810 |
| Abstract: | We monitored Chlamydia trachomatis growth in HeLa cells cultured with either DMEM or RPMI medium containing 10% FCS under 2% or 21% O-2 conditions for 2 days. Bacterial numbers, host cell numbers, and fibrosis related gene expression in the host cells were estimated by an inclusion forming unit assay, a cell counting assay, and a PCR array, respectively. In contrast to RPMI, bacterial growth under low oxygen conditions in DMEM rapidly decreased with increasing host cell density. The addition of supplements (glucose, glutamine, vitamin B12, D-biotin, non-essential amino acids, glutathione) to the media had no effect. The growth of host cells in DMEM under low oxygen conditions rapidly decreased, although the cells remained healthy morphologically. Furthermore, the downregulation of 17 genes was observed under low oxygen in DMEM. Whereas no effect on bacterial growth was observed when culturing in RPMI medium at low oxygen, and the downregulation of three genes (CTGF, SERPINEI, JUN) was observed following bacterial infection compared with the uninfected control cells. Thus, our findings indicate the need for carefully selected culture conditions when performing experiments with C. trachomatis under low-oxygen environments, and RPMI (rather than DMEM) is recommended when a low host cell density is to be used, proposing the major modification of cell culturing method of C. trachomatis in a low-oxygen environment. |
| Rights: | © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ | | http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/75976 |
| Appears in Collections: | 保健科学院・保健科学研究院 (Graduate School of Health Sciences / Faculty of Health Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 山口 博之
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