Liquid culture of N. meningitidis - advice needed! (Jan/04/2011 )

Hi,

I want to grow some N. meningitidis strains in liquid culture so I can harvest the cell culture supernanants for use in other experiments. I have been doing some reading and it looks like I need to culture them in Mueller Hinton broth but I am worried about how to do it safely.

Can anyone advise me on the best way to do this? I know when working with N.m suspension you need to work in a class 2 safety cabinet but I am worried about the incubation steps and also reading the ODs.

Any advice would be greatly appreciated!

We do our neisseria broth culture in a class 3 lab rather than a class 2! But I am not sure that is entirely neccecery or just a bit of overkill on our part.

For incubation just make sure to seal lids firmly and spray with 70% ethanol before taking them out of the hood. For measuring OD's we often put the diluted culture into the cuvette, cover the top with Parafilm and then spray wth 70% ethanol again to ensure no live neisseria left to escape, then leave the parafilm on while taking the OD measurement.