QUESTION about Liebovitz L15 medium

[R Keith Duncan]

I'm a graduate student at the University of Pennsylvania working with 
isolated hair cells from the chick cochlea.  I currently do various 
micromechanics studies on these cells in Liebovitz L15 culture medium 
(with nothing added, i.e. serum, antibiotics) at room temperature.  In 
those conditions we are able to keep the cells viable for 3 hours after
animal sacrifice.  

Recently I have tried warming the cells in culture up to 35 to 40 degrees 
C with a heater that warms the tissue well.  My cells swell and die 
immediately.  As far as I can determine...there is a slight pH change (to 
7.0-7.1) AND there are small bubbles that begin to form around the edges 
of the dish.  Of course there is also an increase in the number of 
bacteria after prolonged heating.  But as I understand it (which is a 
very limited understanding as I'm an engineer), the pH change, bubbles, 
and bacteria would not cause such rapid cell death.  L15 is supposed to 
be stable up to 37 degrees C and is buffered so as to equalize with the 
atmosphere (i.e. I'm not sure if there would be any change in CO2 in the 
medium).

Anyway...I have tried many other things, but nothing has worked.  If any 
of you have experience with this medium or have any suggestions please 
post or email me directly (preferred).  Thanks.

--Keith Duncan
--Univ. of Pennsylvania
--Auditory Research