ECACC Cell Lines are key in vitro tools for SARS-CoV-2 research
Vero E6 (ECACC 85020206): The standard cell line for producing viral stocks of SARS-CoV-2. Yellow: Cell Nuclei, Magenta: Actin Cytoskeleton, Cyan: Mitochondria. (Image courtesy of Ellie Drinkwater, NCPV).
SARS-CoV-2 pandemic response research requires effective in vitro models to help scientists respond to urgent biological questions and to act as potential production platforms for vaccines. Soon after SARS-CoV-2 was identified as the causative agent of COVID-19 it became evident that the virus did not replicate in either embryonated hens’ eggs nor in MDCK cells that are commonly used to isolate and produce influenza vaccines(1) (2). Similarly, some cell lines of the cell lines used previously to cultivate and isolate SARS-CoV-1 in the early 2000s(3) such as MV1Lu (ECACC 88050503) and CV-1 (ECACC 87032605) proved to be less effective hosts for SARS-CoV-2(2).
Angiotensin-converting enzyme-2 (ACE2) has been recognised as the primary receptor of the virus(4) together with priming of the virus spike protein by cellular transmembrane serine protease 2 (TMPRSS2)(5). Since SARS-CoV-2 was discovered in early 2020 there have been extensive efforts to identify suitable human and animal cell lines, particularly those that express ACE2 and TMPRSS2 for use in vitro research into the disease. The African Green Monkey Cell Line ‘Vero E6’ (also known as Vero C1008, Vero 76 or clone E6) (ECACC 85020206), deficient in α and β interferon expression, is now generally accepted as being the standard cell line for the production of viral stocks of SARS-Cov-2 and as a platform for plaque assays(6). Table 1 summarises SARS-CoV-2 susceptible cell lines available from ECACC.
|
Cell Line Name |
ECACC Number |
Species |
*CPE? |
Comment |
Reference |
|---|---|---|---|---|---|
|
BGM |
90092601 |
Green Monkey |
Yes |
|
Wurtz et al(7) |
|
Vero/hSLAM |
04091501 |
Green Monkey |
Yes |
Transfected with lymphocytic activation molecule (SLAM) CDw150 (Measles Virus Receptor)(8) |
Wurtz et al(7) |
|
MA104 |
85102918 |
Green Monkey |
Yes |
|
Wurtz et al(7) |
|
Vero |
84113001 |
Green Monkey |
Yes |
Also known as Vero 81 (ATCC CCL 81) |
Wurtz et al(7) Wang et al(2) |
|
Vero C1008 |
85020206 |
Green Monkey |
Yes |
Also known as Vero E6, Vero 76 or clone E6. High gene and protein expression of ACE2(2) |
Wurtz et al(7) Wang et al(2) |
|
LLC MK2 (Original) |
85062804 |
Rhesus Monkey |
No |
CPE not observed(7) |
Wurtz et al(7) |
|
CACO-2 |
86010202 |
Human Colon |
No |
Mottled appearance of infected monolayer(7) |
Wurtz et al(7) |
|
CRFK |
86093002 |
Cat Kidney |
No |
May support a low level of infection. High gene expression of ACE2(2) |
Wang et al(2) |
|
RK13 |
00021715 |
Rabbit Kidney |
No |
Increased viral load as assessed by viral genome copy number |
Chu et al(9) |
|
Huh-7D12 |
01042712 |
Human Liver |
With lab strain(10) |
Serial passaging of SARS-CoV-2 generated an adapted strain able to infect and induce CPE in Huh-7 cells. Cells not susceptible to clinical isolates(10). |
Ramirez et al(10) |
|
A549 |
86012804 |
Human Lung |
No |
Viral replication of Huh-7 adapted lab strain. Cells not susceptible to clinical isolates(10). |
Ramirez et al(10) |
|
CALU 1 |
93120818 |
Human Lung |
No |
Viral replication of Huh-7 adapted lab strain. Cells not susceptible to clinical isolates(10). |
Ramirez et al(10) |
|
293-T |
12022001 |
Human Kidney |
No |
ACE2 Transfected 293-T cells supported SARS-CoV-2 expression and used to demonstrate importance of TMPRSS2 involvement(5) |
Hoffmann et al(5) |
|
BHK-21 |
ECACC has several BHK-21 subclones including 85011433 (BHK21 (clone 13)) |
Hamster Kidney |
No |
ACE2 Transfected BHK-21 cells supported SARS-CoV-2 expression and used to demonstrate importance of TMPRSS2 involvement(5) |
Hoffmann et al(5) |
|
SH-SY5Y |
94030304 |
Human Neuroblastoma |
No |
SARS-CoV-2 modestly invaded cells with no obvious CPE. Retinoic Acid (RA) differentiation upregulated ACE2 and TMPRSS2 expression(11) |
Bielarz(11) |
|
SK-N-BE(2) |
95011815 |
Human Neural Tumour |
No |
SARS-CoV-2 modestly invaded cells with no obvious CPE. Retinoic Acid (RA) differentiation upregulated ACE2 and TMPRSS2 expression(11) |
Bielarz(11) |
|
U-87 MG |
89081402 |
Human |
No |
SARS-CoV-2 modestly invaded cells with no obvious CPE. Retinoic Acid (RA) differentiation upregulated ACE2 and TMPRSS2 expression(11) |
Bielarz(11) |
|
U-373 MG (Uppsala) |
08061901 |
Human |
No |
SARS-CoV-2 modestly invaded cells with no obvious CPE. Retinoic Acid (RA) differentiation upregulated ACE2 and TMPRSS2 expression(11) |
Bielarz(11) |
Table 1. SARS-Cov-2 susceptible cell lines available from ECACC.
*CPE = Cytopathic Effect
References
1. Barr IG, Rynehart C, Whitney P, Druce J. SARS-CoV-2 does not replicate in embryonated hen’s eggs or in MDCK cell lines. Eurosurveillance. 2020 Jun 25;2525:2001122.
2. Wang L, Fan X, Bonenfant G, Cui D, Hossain J, Jiang N, et al. Susceptibility to SARS-CoV-2 of Cell Lines and Substrates Commonly Used to Diagnose and Isolate Influenza and Other Viruses. Emerg Infect Dis. 2021 May;275:1380–92.
3. Kaye M, Druce J, Tran T, Kostecki R, Chibo D, Morris J, et al. SARS–associated Coronavirus Replication in Cell Lines. Emerg Infect Dis. 2006 Jan;121:128–33.
4. Bourgonje AR, Abdulle AE, Timens W, Hillebrands J, Navis GJ, Gordijn SJ, et al. Angiotensin‐converting enzyme‐2 (ACE2), SARS‐CoV‐2 and pathophysiology of coronavirus disease 2019 (COVID‐19). J Pathol [Internet]. 2020 May 17 [cited 2021 Dec 1]; Available from here.
5. Hoffmann M, et al. SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell. 2020 Apr 16;1812:271–280.e8.
6. Cagno V. SARS-CoV-2 cellular tropism. Lancet Microbe. 2020 May 1;11:e2–3.
7. Wurtz N, Penant G, Jardot P, Duclos N, La Scola B. Culture of SARS-CoV-2 in a panel of laboratory cell lines, permissivity, and differences in growth profile. Eur J Clin Microbiol Infect Dis. 2021 Mar 1;403:477–84.
8. Ono N, Tatsuo H, Hidaka Y, Aoki T, Minagawa H, Yanagi Y. Measles Viruses on Throat Swabs from Measles Patients Use Signaling Lymphocytic Activation Molecule (CDw150) but Not CD46 as a Cellular Receptor. J Virol [Internet]. 2001 May 1 [cited 2021 Dec 1]; Available from here.
9. Chu H, et al. Comparative tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV with implications for clinical manifestations, transmissibility, and laboratory studies of COVID-19: an observational study. Lancet Microbe. 2020 May 1;11:e14–23.
10. Ramirez S, Fernandez-Antunez C, Galli A, Underwood A, Pham LV, Ryberg LA, et al. Overcoming Culture Restriction for SARS-CoV-2 in Human Cells Facilitates the Screening of Compounds Inhibiting Viral Replication. Antimicrob Agents Chemother [Internet]. 2021 Apr 26 [cited 2021 Nov 30]; Available from here.
11. Bielarz V, et al. Susceptibility of neuroblastoma and glioblastoma cell lines to SARS-CoV-2 infection. Brain Res. 2021 May 1;1758:147344.
Written by Jim Cooper PhD, December 2021