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Next Subject 		How do I grow LL2 (murine Lewis Lung)LadyJustin2009-12-03Click here to register.
I need to start growing Lewis Lung cells for an in-vivo study. One of the researchers has a vial of frozen cells to give me so it's not coming with instructions on how to grow. What media do I use? I'm assuming this is an adhesion cell line? What can I expect form growth rate? How often will it need to be split? What problems might I encounter?
 
Re: How do I grow LL2 (murine Lewis Lung)
lubrol2009-12-11Click here to register.
A derivative of this tumor is available from the ATCC as ATCC® Number: CRL-1642™. The specifications there recommend DMEM plus 10% FBS for a complete growth medium.

The cells have a doubling time of around 21 hours at 37 C and grow as a mixed population of adherent and suspension cells. This latter point can cause some issues depending on your needs. But it also is confusing in that the culture looks either contaminated or unhealthy even when it is fine.

The cells need to be split 1:3 to 1:6 when subcultured: the exact ratio will depend on how well they are growing and what your needs are in terms of cell density and timing. Being a cancer line, they will be fairly easy to grow.

You may be able to "purge" the non-adherent cells by frequent and thorough washing with PBS prior to re-feeding; but I suspect that that might change the properties of the culture.


Hope this helps,

Lubrol

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