Invivogen
Menu

Human ACE2 & TMPRSS2 expressing A549 Cells

A549-hACE2-TMPRSS2 Cells Unit size Cat. code Docs Qty Price
A549 lung carcinoma cells expressing human ACE2 and human TMPRSS2
3-7 x 10e6 cells
a549-hace2tpsa
+-
$1,374.00
A549-hACE2 Cells Unit size Cat. code Docs Qty Price
A549 lung carcinoma cells expressing human ACE2
3-7 x 10e6 cells
a549-hace2
+-
$1,260.00

You may also need : Spike-expressing vectors for lentiviral pseudotyping | View more associated products

A549 lung carcinoma cells expressing the SARS-CoV-2 receptors ACE2 and TMPRSS2

Permissivity of A549-hACE2 and A549-hACE2-TMPRSS2 cells to SARS-CoV-2 infection
Permissivity of A549-hACE2 and A549-hACE2-TMPRSS2 cells to SARS-CoV-2 infection

 

InvivoGen offers A549-hACE2-TMPRSS2 cells and A549-hACE2 cells which are derived from the human A549 lung carcinoma cell line. The A549 cell line is a commonly used model for the study of respiratory infections.
SARS-CoV-2 (2019-nCoV), the causative agent of coronavirus disease-19 (COVID-19), gains entry into the human lung epithelium (ciliated, club, and basal cells) through the interaction of ACE2 and TMPRSS2 receptors with the virus Spike protein [1, 2].

A549 wild-type cells are poorly permissive to infection by pseudotyped lentiviral particles expressing the SARS-CoV-2 Spike protein, as they express negligible levels of ACE2 and no TMPRSS2 [2, 3, in-house data].

  • ACE2 and ACE2-TMPRSS2 A549 cells are permissive to infection by pseudotyped lentiviral particles expressing the SARS-CoV-2 Spike protein.
  • The addition of TMPRSS2 expression significantly increases the cell line infectivity.
     

Features of A549-hACE2 (and TMPRSS2) Cells:

  • Verified overexpression  of human ACE2 by RT-qPCR, and ACE2 cell surface expression by FACS using a Spike-S1-Fc fusion protein
  • Verified overexpression of human TMPRSS2 by RT-qPCR
  • Functionally-tested in infection assays with pseudotyped lentiviral particles expressing the SARS-CoV-2 Spike protein
     

Applications of A549-hACE2 (and TMPRSS2) Cells:

  • Screening of small molecule inhibitors and/or neutralizing antibodies of the ACE2‑Spike interaction
  • Screening of small molecule inhibitors and/or neutralizing antibodies of the TMPRSS2 surface protease
  • Comparative studies of the effects of drugs targeting ACE2 and/or TMPRSS2 on the SARS-CoV-2 infection and cellular signaling outcomes
     

ACE2 and TMPRSS2 background:

ACE2 (angiotensin I-converting enzyme-2) and TMPRSS2 (transmembrane protease serine 2)play a critical role in the pathogenesis of COVID-19 by allowing viral entry into target cells (e.g. human lung epithelium). ACE2 and TMPRSS2 are cell-surface proteins that both interact with the virus Spike (S) protein [1-3]. ACE2 is mandatory for the binding of SARS-CoV-2 at the cell surface through its interaction with the Spike receptor-binding domain (RBD) [4]. Following this, TMPRSS2 cleaves the S protein into two functional subunits (S1 and S2), allowing virus-host membrane fusion, and the release of viral contents (e.g. RNA) into the cytosol [3-5]. Another protease, the Cathepsin L, also mediates cleavage of the S protein but it acts in the endosomes. Camostat is a clinically-proven inhibitor of TMPRSS2 and has been shown to inhibit SARS-CoV-2-S pseudotyped viral particles entry into primary human lung cells in a dose-dependent manner [2]. This observation demonstrates the critical implication of TMPRSS2 in SARS-CoV-2 infection and spread. Moreover, in lung cells that fail to express robust levels of Cathepsin L, the virus entry depends on a furin-mediated pre-cleavage of the S protein at the S1/S2 site, before subsequent TMPRSS2-mediated cleavage at the S2' site [6].

 

Learn more on SARS-CoV-2Learn more about SARS-CoV-2 infection cycle, immune responses, and potential therapeutics.

 

References

1. Chen H. et al., 2020. SARS-CoV-2 activated lung epithelia cell proinflammatory signaling and leads to immune dysregulation in COVID-19 patients by single-cell sequencing. medRxiv: DOI 10.1101/2020.05.08.20096024.
2. Hoffmann M. et al., 2020. SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell. 181:1-16.
3. Matsuyama S. et al., 2020. Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells. PNAS. 117(13):7001-7003.
4. Zhou P. et al., 2020. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 579(7798):270-273.
5. Walls A.C. et al., 2020. Structure, function, and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell. 181(2):281-292.e6.
6. Hoffman M. et al., 2020. A multibasic cleavage site in the Spike protein of SARS-CoV-2 is essential for infection of human lung cells. Molecular Cell. 78:;1-6.

Figures

Infection of A549-hACE2-TMPRSS2 cells by SARS-CoV-2 Spike pseudotyped lentiviral particles
Infection of A549-hACE2-TMPRSS2 cells by SARS-CoV-2 Spike pseudotyped lentiviral particles

Infection of A549-hACE2-TMPRSS2 cells by Spike (D614 or G614) pseudotyped lentiviral particles.

~2x104 A549-hACE2-TMPRSS2 cells were cultured in the presence of SARS-CoV-2 Spike D614-variant (up panels) or Spike G614-variant (bottom panels) pseudotyped GFP lentiviral particles. After 72h, the transduction efficiency of the Spike pseudotyped GFP particles was evaluated by fluorescence microscopy.

Validation of ACE2 surface expression on A549-hACE2-TMPRSS2 cells by FACS
Validation of ACE2 surface expression on A549-hACE2-TMPRSS2 cells by FACS

Surface expression of hACE2 by A549-hACE2-TMPRSS2 cells.
~2x105 A549 (wild-type) and A549-hACE2-TMPRSS2 cells were incubated with 1 μg of Spike-S1-Fc or CTLA-4-Fc fusion proteins for 1h at 4°C. Cells were then washed and incubated with 0.5 μg of a goat anti-hIgG1-Fc antibody coupled to PE for 1h at 4°C. Cell surface staining was analyzed by flow-cytometry.

Validation of ACE2 and TMPRSS2 overexpression by RT-qPCR
Validation of ACE2 and TMPRSS2 overexpression by RT-qPCR

Human ACE2 and TMPRSS2 mRNA expression in A549-hACE2-TMPRSS2 cells.

Total mRNA was extracted from ~1x106 A549 (wild-type) and A549-hACE2-TMPRSS2 cells. ACE2 and TMPRSS2 mRNA were amplified using quantitative RT-qPCR. Data are represented as the log2 fold change comparing ACE2 or TMPRSS2 expression to a housekeeping gene. ND: not detected.

Infection of A549-hACE2 cells by SARS-CoV-2 Spike pseudotyped lentiviral particles
Infection of A549-hACE2 cells by SARS-CoV-2 Spike pseudotyped lentiviral particles

Infection of A549-hACE2 cells by Spike (D614 or G614) pseudotyped lentiviral particles.

~2x104 A549-hACE2 cells were cultured in the presence of SARS-CoV-2 Spike D614-variant (up panels) or Spike G614-variant (bottom panels) pseudotyped GFP lentiviral particles.

After 72h, the transduction efficiency of the Spike pseudotyped GFP particles was evaluated by fluorescence microscopy.

Validation of ACE2 surface expression on A549-hACE2 cells by FACS
Validation of ACE2 surface expression on A549-hACE2 cells by FACS

Surface expression of hACE2 by A549-hACE2 cells.

~2x105 A549 (wild-type) and A549-hACE2 cells were incubated with 1 μg of Spike-S1-Fc or CTLA-4-Fc fusion proteins for 1h at 4°C.
Cells were then washed and incubated with 0.5 μg of a goat anti-hIgG1-Fc antibody coupled to PE for 1h at 4°C. Cell surface staining was analyzed by flow-cytometry.

Validation of ACE2 overexpression by RT-qPCR
Validation of ACE2 overexpression by RT-qPCR

Human ACE2 mRNA expression in A549-hACE2 cells.

Total mRNA was extracted from ~1x106 A549 (wild-type) and A549-hACE2 cells. ACE2 mRNA was amplified using quantitative RT-qPCR.
Data are represented as the log2 fold change comparing hACE2 expression to a housekeeping gene. ND: not detected.

Back to the top

Specifications

Growth medium: DMEM, 4.5 g/L glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum (FBS), 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™
 

Antibiotic resistance:

Quality Control:

  • ACE2 gene expression has been verified by RT-qPCR, FACS staining, and functional assays.
  • TMPRSS2 gene expression has been verified by RT-qPCR, and functional assays.
  • The stability for 20 passages, following thawing, has been verified for A549-hACE2 and A549-hACE2-TMPRSS2 cells.
  • These cells are guaranteed mycoplasma-free. 
     

InvivoGen's products are covered by a Limited Use License (See Terms and Conditions).

Back to the top

Contents

Please note: Each cell line is sold separately. See TDS for the exact contents of each cell line. 

A549-hACE2 cells

  • 3-7 x 106 A549-hACE2 cells in a cryovial or shipping flask
  • 1 ml of Puromycin (10 mg/ml)
  • 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.

A549-hACE2-TMPRSS2 cells

  • 3-7 x 106 A549-hACE2-TMPRSS2 cells in a cryovial or shipping flask
  • 1 ml of Puromycin (10 mg/ml)
  • 1 ml of Hygromycin (100 mg/ml)
  • 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.
     

Shipped on dry ice (Europe, USA & Canada)

 

Back to the top

FAQ

Visit our FAQ Any questions about our cell lines ? Visit our frequently asked questions page

Back to the top
Customer Service
& Technical Support
Shopping cart is empty