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Human ACE2 Stable Cell Line - HEK293T

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Cat. No. Product Name Transgene Description Price
CSC-ACE01 Human HEK293T (ACE2) Stable Cell Line Human ACE2 One frozen vial containing millions of cells. Inquiry
CSC-ACE02 Human HEK293T (hACE2-hTMPRSS2) Stable Cell Line Human ACE2
Human TMPRSS2
One frozen vial containing millions of cells. Inquiry

CELL LINE INFORMATION

Cell Line Description
This cell line is constructed by transduction of human angiotensin I converting enzyme 2 (ACE2) into HEK293T cells, followed by stable cell selection. HEK293T is derived from HEK293 and is commonly used in scientific research. HEK293T-human ACE2 cell line can be used for in vitro screening and characterization of drug candidates against SARS-CoV, SARS-CoV-2 (2019-nCoV).
Background

ACE2 protein belongs to the angiotensin-converting enzyme family of dipeptidyl carboxydipeptidases and has considerable homology to human angiotensin 1 converting enzyme. This protein catalyzes the cleavage of angiotensin I into angiotensin 1-9, and angiotensin II into the vasodilator angiotensin 1-7. The organ- and cell-specific expression of this gene suggests that it may play a role in the regulation of cardiovascular and renal function, as well as fertility. In addition, this protein is a functional receptor for the spike glycoprotein of the human coronavirus HCoV-NL63 and the human severe acute respiratory syndrome coronaviruses, SARS-CoV and SARS-CoV-2 (COVID-19 virus).

hTMPRSS2 protein belongs to the serine protease family. The encoded protein contains a type II transmembrane domain, a receptor class A domain, a scavenger receptor cysteine-rich domain and a protease domain. Serine proteases are known to be involved in many physiological and pathological processes. This gene was demonstrated to be up-regulated by androgenic hormones in prostate cancer cells and down-regulated in androgen-independent prostate cancer tissue. The protease domain of this protein is thought to be cleaved and secreted into cell media after autocleavage. This protein also facilitates entry of viruses into host cells by proteolytically cleaving and activating viral envelope glycoproteins. Viruses found to use this protein for cell entry include Influenza virus and the human coronaviruses HCoV-229E, MERS-CoV, SARS-CoV and SARS-CoV-2 (COVID-19 virus).

Host Cell
HEK293T

SAFETY AND PACKAGING

Mycoplasma
Negative
Format
One frozen vial containing millions of cells
Storage
Liquid nitrogen
Safety Considerations

The following safety precautions should be observed.

  1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.
  2. No eating, drinking or smoking while handling the stable line.
  3. Wash hands after handling the stable line and before leaving the lab.
  4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.
  5. All waste should be considered hazardous.
  6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship
Dry ice

REPRESENTATIVE DATA

Figure 1. Cell morphology of HEK293T-ACE2 cell line under bright field (left) and GFP fluorescence (right).

Figure 2. Infection assay. 72 h after infection of 20 μL SARS-CoV-2 S glycoprotein pseudotyped GFP lentivirus into HEK293T-ACE2 cells, under bright field (left) and GFP fluorescence (right).

Figure 3. qPCR analysis of human ACE2 expression in HEK293T-ACE2, HEK293T is used as control.

Figure 4. Western blot analysis of human ACE2 expression in HEK293T-ACE2, HEK293T is used as control.

Q & A

Q: Does this cell line express full length protein on cell surface (805 amino acid is full length)?

A: Yes, it express full length human ACE2 protein.

Q: Did FACS analysis was done to ensure the protein is present on cell surface?

A: No. But our ACE2/HEK293T can be successfully infected by Pseudotyped GFP rSARS-CoV-2 Spike (CD Cat# COV-PS02) which means cells contain functional ACE2.

Q: Do CSC-ACE01 cell line require a selection antibiotic in the media? if so, which one?

A: We use 1 μg/mL of puromycin.

Q: How do you transduce ACE2 and TMPRSS2 into this cell line (CD Cat# CSC-ACE02)?

A: The cells were transduced using a second generation lentiviral vector carrying a transgene for hsACE2 (Accession# NM_021804.3) and hTMPRSS2 (Accession# NM_001135099.1), Integration and expression were selected for by incubation with Puromycin and Blasticidin.

Q: For CSC-ACE01, I was wondering how many times the ACE2 cell line could be passaged before the ACE2 receptors started to fall off?

A: Maintain the cells in Puromycin to ensure the ACE2 transgene is expressing. When grown under selection, the receptor levels should not decrease with passage. We have carried the ACE2 cell line to P30 without losing transduction efficiency.

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