Elwood Siagian

Seethis site. US CDC has grows them in cell culture. And deposit at BEI (see the site). It is free to get the isolate, but you have to pay a fee. You can find all the info in Q & A sections. There is an email, probably you can ask from there. Be ware, there are different strains out there already (due to mutation). You need to find out which strain you like to study.

CDC Grows SARS-CoV-2, the virus that causes COVID-19, in Cell Culture

Virus isolates will be used for research by the scientific and medical community

https://www.cdc.gov/coronavirus/2019-ncov/php/grows-virus-cell-culture.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.gov%2Fcoronavirus%2F2019-ncov%2Fabout%2Fgrows-virus-cell-culture.html

Dear Elwood

the following paper would be helpful:

dx.doi.org/10.17504/protocols.io.bcduis6w

Since the virus has has made the lung its target, I will suggest the use of human lung cell line. Human normal lung fibroblasts HEL-299 and MRC-5 cells are some examples.

This may help you with some idea:

An alternative workflow for molecular detection of SARS-CoV-2 - escape from the NA extraction kit-shortage, Copenhagen, Denmark, March 2020. Euro Surveill. 2020 Apr 09;25(14).

https://doi.org/10.2807/1560-7917.ES.2020.25.14.2000398

https://www.cdc.gov/coronavirus/2019-ncov/php/grows-virus-cell-culture.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.gov%2Fcoronavirus%2F2019-ncov%2Fabout%2Fgrows-virus-cell-culture.html

Also, you could try commecially available primary endothelial cells such as human lung microvascular endothelial cells:

https://bioscience.lonza.com/lonza_bs/US/en/Primary-and-Stem-Cells/p/000000000000184961/HMVEC-L-%E2%80%93-Human-Lung-Microvascular-Endothelial-Cells

Viro cell lines (VeroE6/TMPRSS2), Caco-2, HEK 293T, Huh7, LLC-MK2

Dear Elewood,

Just read my text. Findings by researcher might be helpful for your query.

One of the excellent work was carried out by Chu and his colleagues publication work (The Lancet Microbe

,Volume 1, Issue 1, May 2020). They identified several cell lines in which both SARS-CoV and SARS-CoV-2 replicated efficiently, but cytopathic effects were only seen in the non-human primate kidney cell lines VeroE6 and FRhK4, including cell rounding, detachment, and degeneration. VeroE6 is the standard cell line used to produce viral stocks of SARS-CoV-2 in different laboratories and to do plaque assays. The noted cytopathic effects are consistent with use of this cell line in virology because of the susceptibility of VeroE6 cells to infection, lower saturation density (optimal for plaque formation), and interferon α and interferon β secretion deficiency.

Studies on Animals-

Further, they noted studied on animals, Chu and colleagues analysed several other animal epithelial kidney cell lines. A significant increase of viral load between 2 h postinoculation (hpi) and 120 hpi was seen in LLCMK2 (monkey), RK-13 (rabbit), PK-15 (pig), and CRFK (cat) cell lines. These results do not fully recapitulate the animal tropism reported so far: findings of studies of SARS-CoV-2 and SARS-CoV have highlighted possible infections in cats (and felines in general), ferrets, and hamsters, whereas rabbits were not investigated, dogs seemed (until now) to be poorly permissive, and pigs were not infectable. These discrepancies might derive from the kidney origin of the cells, their immortalisation, or the culturing conditions.

****Studies on human-

Chu and colleague also compared various human cell lines, including pulmonary (Calu3), intestinal (Caco2), hepatic (Huh7), and neuronal (U251) cells. However, some anatomical sites or cell types (eg, cardiomyocytes) were not represented or only one cell line was used to represent a site; for example, with use of only U251 cells, the presented findings are not sufficient to draw conclusions on neuroinvasiveness, particularly since they contrast with previous research. SARS-CoV-2 replication was greatest in Calu3 and Caco2 cell lines, and these anatomical sites (pulmonary and intestinal) are the ones positive for viral RNA in humans. In their studies, on the other hand, SARS-CoV was shown to replicate better than SARS-CoV-2 in Caco2 cell lines.

*****What they conclude in their work output, the study by Chu and colleagues offers possible directions for designing more in-depth studies about animal and human susceptibility to SARS-CoV-2 infection. However, we should keep in mind that results obtained in cell lines might not always recapitulate the effects in the whole organism.

Refer the site given below for comprehensive purpose.

https://www.thelancet.com/journals/lanmic/article/PIIS2666-5247(20)30008-2/fulltext

Hope this is helpful for you.

Ashish

There are some paper that state that Vero E6 is one of the cell line that exhibit a noticeable morphological changes or cytopathic effect (CPE) compared to Vero CCL81

Our team still works with Vero CCL81 and still exploring and identifying the CPE

Maybe the article entitled Severe Acute Respiratory Syndrome Coronavirus 2 from Patient with Coronavirus Disease, United states by Jennifer Harcourt et.al. in CDC websites could help