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. 2020 Mar 31;117(13):7001-7003.
doi: 10.1073/pnas.2002589117. Epub 2020 Mar 12.

Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

Shutoku Matsuyama  1 , Naganori Nao  2 , Kazuya Shirato  2 , Miyuki Kawase  2 , Shinji Saito  3 , Ikuyo Takayama  3 , Noriyo Nagata  4 , Tsuyoshi Sekizuka  5 , Hiroshi Katoh  2 , Fumihiro Kato  2 , Masafumi Sakata  2 , Maino Tahara  2 , Satoshi Kutsuna  6 , Norio Ohmagari  6 , Makoto Kuroda  5 , Tadaki Suzuki  4 , Tsutomu Kageyama  3 , Makoto Takeda  1
Affiliations

Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

Shutoku Matsuyama et al. Proc Natl Acad Sci U S A. .

Abstract

A novel betacoronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which caused a large respiratory outbreak in Wuhan, China in December 2019, is currently spreading across many countries globally. Here, we show that a TMPRSS2-expressing VeroE6 cell line is highly susceptible to SARS-CoV-2 infection, making it useful for isolating and propagating SARS-CoV-2. Our results reveal that, in common with SARS- and Middle East respiratory syndrome-CoV, SARS-CoV-2 infection is enhanced by TMPRSS2.

Keywords: SARS-CoV-2; TMPRSS2; VeroE6; coronavirus; outbreak.

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Conflict of interest statement

The authors declare no competing interest.

Figures

Fig. 1.
Fig. 1.
(A) Expression of TMPRSS2 in total cellular RNA (0.2 µg) of indicated cells was compared with that in human lung RNA (catalog no. 636524; Clontech) by quantitative real-time PCR. ND, not detectable. (B) SARS-CoV-2–infected VeroE6/TMPRSS2 cells. Cell rounding (black arrows) and syncytium formation (white arrows) (C). Electron micrograph showing isolated virus particles with negative staining. (Scale bar, 200 nm.) (D) Viral RNA multiplication in various cells at 48 h postinoculation with the viral specimen, as determined by real-time RT-PCR using E and N primer/probe sets (9). Cq, quantitation cycle. (E) Real-time RT-PCR amplification plot using the E primer/probe set, corresponding to the data in C. RFU, relative fluorescence units. (F) Comparison of cell susceptibility to the isolated virus, detected with a patient’s serum and Alexa 488-conjugated goat anti-human IgG. Nuclei were stained with DAPI.
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