Problem with Neuronal Culture

Jerome Freed jjfreed at
Mon Aug 25 14:55:38 EST 1997

Bill Marszalec wrote:

>    I would like to pose a problem to those members of this group who 
> have worked with cultured neurons or cultured cells in general.  For the 
> past three years, we have been performing whole cell voltage clamp 
> experiments using a primary culture of isolated frontal cortical neurons 
> prepared from 17 day embryonic rat pups.  My question relates to a 
> seemingly cyclic fluctuation of the quality of these cells.
It would be very prudent to test your cultures (good and bad) for
mycoplasma.  The method of fluorescent staining of 3T3 cell cultures
inoculated with some of the medium from your neuronal cultures, using a
DNA stain like Hoechst or DAPI, would be the simplest to carry out.

Further, is it possible to passage the evil influence (or putative
agent) to some other cultures that seem normal and transfer the
cytopathology you have observed?

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