Dear flowers I have never worked with the cell line WEHI 164 clone 13 and have a couple of questions for those of you that may have worked with them. The cells seem to grow as "spindle-shaped cells" and round cells that adhere and also some in suspension. I am trying to use these cells as targets for TNF quantitation assay (WEHI clone 13 is suppose to be a more sensitive cells line for this purpose). I am not sure if the morphology of these cells is correct or is it something that we are doing wrong. We are using the recomended media (RPMI-1640) and the recomended supplements (10% FCS, etc) as stated in the ATCC attached protocol. Do they ever reach true confluency? We can grow them to about 60% confluency (mixed bag of spindle shaped cells and rounded attached cells) The media becomes acidic overnight at this stage. We are currently talking to ATCC and trying to get some information from them, but I thought I asked this group to determine if anyone had experience in this area. Any experienced users out there that can shed some light on this? Thanks Ed Cabrera, PhD. Procter and Gamble Pharmaceuticals Mason, OH
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