Culture of lymphocytes

[Jorg Kirberg]

In article <3gatm7$c3j at lynx.unm.edu>, inkim at carina.unm.edu (In C. Kim) wrote:

> Cell culture is performed generally in a humidified, CO2-incubator.  In
> order to prepare metaphase spreads, lymphoyctes are cultured for only
> three days in the presence of a mitogen and colicimide.  I wonder if this
> condition requires CO2 or not.  Could anybody have experience in culturing
> human peripheral lymphocytes for genetic research? 
> 


Dear inkim,

CO2 (and NaHCO3 in the culture medium) are needed to get a high buffering
capacity. You might try without (replacing the NaHCO3 with NaCl) in the
absence of CO2, but as you use highly activated cells I dont know if the
resulting medium is buffered enough to keep the pH in a physiological
range. Adding more HEPES might help in that case.
Maybe just try and look at the medium afterwards. If it is totally yellow
forget about it.
(Actually if you just leave out the CO2 it get's pink; that pH is toxic as
well)

jorg

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Joerg Kirberg                     EMAIL: kirberg at bii.ch
Basel Institute for Immunology    FAX:   41-61-605 13 72
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