Dear fellow scientists,
Has anyone had success in culturing and isolating SARS Cov-2 viruses?
We and our team still currently working on culturing in aim to isolate the virus, we are using Vero Cell CCL-81 and has already performed several blind passages. But we are still struggling to identify the CPE caused by SARS Cov-2 infection as it appears to exhibit a non-specific pattern.
May we ask fellow scientists that working in similar projects to share their knowledge according to this issue?
Best Regards,
Andi YW
Faculty of Medicine, Airlangga University, Surabaya, Indonesia,
I would just like to start with the first one, published today in Neurology, on the cytopathic effects of the virus on the brain. A lot more research is required on the subject, but this article does mention your codeword.
Published Online:Jun 16 2020https://doi.org/10.1148/radiol.2020202222
https://www.researchgate.net/publication/341654297_Covid-19_A_Silent_Killer
This paper help you out
https://www.researchgate.net/publication/341654297_Covid-19_A_Silent_Killer
This paper help you out
Pardon, but what we would like to ask is cell culture morphological changes.
Hi all participants and have a nice week. In the following RG open question I attempt to search causes of the Lombardia hyper cluster of COVID19,
https://www.researchgate.net/post/The_novel_Coronavirus_in_N_Italy_Lombardia_COVID19_2019nCoV_SARSCoV2_shows_a_fatality_rate_compatible_with_SARS-MERS_Why
a very worst case.
In order to obtain data, there are extensive bibliographical lists about
--oral-fecal contagion
--chemicals vs. SARS-CoV/2
--receptor ecosystem
--animal reservoirs
--Guillain-Barrè syndrome
-- co-infection (other local CoV)
-- comparative data vs. others world hyper clusters
I hope this is an interesting point to obtain data for this dramatic situation in N. Italy (and Central Europe), in order to prevent similar evolution in the next future.
-sv-
Cytipathogenic Effect - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1829448/
Isolation of Viral Culture - Article SARS-CoV-2 infection diagnosed only by cell culture isolatio...
Hello,
We do a lot of SARS-CoV2 work at our lab and we use Vero-E6 cells (ATCC® CRL-1586™, clone E6) for this purpose which has good amount of compatible ACE2 expression to support viral growth. These cells should be maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS), penicillin-streptomycin, MEM non-essential amino-acids (Gibco; 25025CL) and Hepes buffer (Gibco; 15630-080) at 37°C with 5% CO2. And Of course cells free of mycoplasma is the ideal way to go. This should work!
Good luck
Dear Raveen Rathnasinghe,
Thank you for your clear information about culture methods
But may we ask about the morphological changes that are known to act as SARS-COV-2 cytopathic effect or viral infection of the cells?
Best Regards,
Andi YW
Faculty of Medicine, Airlangga University
In the above mentioned cells, we see the "typical" cytopathic effect. Theses viruses typically get peak titers after 48 hours since infection.
What you should see is,
1) detachment of cells
2) Elongated morphology
3) Syncytium formation (not always)
If your viruses are viable and the cells susceptible for SARS-CoV2 infection, you should be able to see CPE without adding any exogenous trypsins. I would recommend infecting confluent monolayers with different MOIs to establish your infection parameters . I have attached a picture for your reference. This an infection done using SARS-CoV2 (USA-WA1/202 isolate) in Vero-E6 cells with a very low MOI. These monolayers were fixed using 4% PFA so that is why you don't see any cells floating.
