We at our lab have established the serological and molecular diagnosis of various viruses. Now we are in a try to establish the viral culture facility at our lab. it would be really great if anyone can help us by sharing information about the basic needs like consumables, reagents and available ready made media and reagents for the same. we have a -80oC, -20oC, BSC B2, CO2 Incubator, Fluorescence Microscope, Inverted Microscope in our lab. Providing any literature for the same will be a great help. Thank you.
You need animal tissue culture facility for virus cultivation
https://www.vanderbilt.edu/viibre/CellCultureBasicsEU.pdf
Hi Ayan,
You will need at least two BSC II cabinets (one for clean cell culture and one for virus cell culture) although you do not have to have them in the same laboratory. Similarly, you will need at least two CO2 incubators (one for clean cells, and one for virus-infected cells) although again you can host the clean cell culture work separately from the virus work ie in a different lab. Many institutes have both clean and virus work co-located in the same room in which case you need good lab practices to ensure you do not infect clean cells with virus inadvertently (eg host clean/virus work on different sides of the lab, different lab coats/gowns for working with virus, preferably different colour too, 0% tolerance for anything going from a virus cabinet/incubator into the clean cabinet/incubator etc).
You will also need a large refrigerator, some -20C freezer space, a water bath, a centrifuge capable of spinning 15mL and 15mL Falcons (preferably refrigerated), a microscopy, a haemocytometer and access to a -80C freezer. You'll need lots of pipettes (preferably dedicated to specific cabinets), a few pipette-boys, and most basic lab equipment (microfuge, vortex, plate rocker etc). Most labs need access to a bench top autoclave as making up certain reagents and sterilising in-house is cheaper than buying in sterile directly
Unfortunately it would take hours to list reagents (and would depend on the cell line), but you will burn through cell culture flasks, cell culture plates, tips, stripettes and Falcon tubes (none of which are cheap). Then there's FCS, media and other additives which are eye-wateringly expensive!
One last word of advice - a BSC B2 is not designed for cell culture / virus culture. You need A2 cabinets (https://www.nuaire.com/how-class-two-bsc-work) as best practice requires a curtain of HEPA-filtered recirculated air passed over the working area in order to protect your sensitive work. If you do not have this curtain of clean air, you will experience far more contamination which will cost you excess time and money!
Lastly, the link posted by Abdul above is an excellent guide on how to perform cell culture and is worth reading in full!
Thank you very much Abdul and Barry. It was really helpful. Will keep in mind your suggestions.
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