I've been culturing LNCaP cells for about a month now, and have realized that after the second or third passage, the cell adherence drops drastically. The floating cells are not dead, and the ones that do adhere are healthy, but fewer and fewer cells attach with each passage. Has anyone else had this problem?
I'm using trypLE instead of trypsin and I keep the dissociation time to a minimum when passaging.
I would check for possible bacterial contamination, though. I've seen this kind of behavior by contaminated cell lines in the past.
During passage, if you are not able to detach all "adherent" cells and you enrich population of loosely attached cells or floating cells, it is likely that you are "selecting" a population of cells that have alterations in their ability to adhere cell culture plates.
I am facing the same problem. By passage 18, the adhering properties have drastically dropped, though the LNCAP cells are 100% live when checked with cytometer and no mycoplasma contamination found. Thawing a new batch now. But, unable to understand the reason behind it. Has it drifted from being "itself" ?
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