616 Chairman: H. CHRISTENSEN 
GurRoFF: What about cooling to 0°? Has that been done with ascites cells? 
CHRISTENSEN: Yes. It is possible to wash the cells very quickly at ice temperatures without loss. 
Letting them stand in such a solution leads to the loss of the gradients. 
E. Rosperts: The total amino acid pool of ascites cells incubated in ascitic fluid 2m vityo for rather 
prolonged periods of time without any added glucose, is maintained very tenaciously. However, 
when we tried to incubate these cells in a variety of buffers, even for brief periods, we found con- 
siderable leakage into the fluid of the easily extractable ninhydrin-reactive constituents. We have 
not yet succeeded in finding in vitro conditions other than incubation in the protein-rich ascitic 
fluid, under which we could maintain the pool as we see them on chromatograms. This finding has 
puzzled me in terms of what it might mean in relation to the various kinetic experiments that have 
been done in buffers rather than in the ascitic fluid itself. 
