using culture sup of one cell line  to induce 2nd cell line - (Jan/28/2002 )

Hi,

Does anyone know of a good protocol that describes how to use  the culture  supernatant (conc. etc)  of one cell line to induce a 2nd cell line.
Any help is appreciated.

We routinely use cultered media to treat other cell lines.  In our procedure, the media is taken from one cell line (cultured media) and stored with the addition of protease inhibitor at -70C until we are ready to use.  One to two days before we want to use the CM, we thaw overnight in the refrigerator then use centriprep tubes to concentrate the sample to 10X.  We then filter sterilize the sample using syringe filters and add to the new cell line at a 1X concentration.  Hope this helps.