I'm growing K562 cells in RPMI media with 10% FBS or 10% FBEssence to check for any growth preference. Cells are growing at the same rate in both, but in FBEssence there is reproducable clumping of cells that seems to be caused by something in the FBEssence (cells swapped from FBS --> FBEssence become clumpy, cells switched from FBEssence --> FBS stop clumping). What should I start to investigate to show a possible difference in cell morphology between the two conditions? Gene expression levels? Cytoskeleton changes? etc ...
Hi,
Monocytes start clumping when they are not 100% happy or sometimes also when they are happy and divide fast. They start to clump when there is very small concentration of cells, very big concentration of cells, too little FBS or Glutamine, infection, change of ph...
If the clumping is the only difference you observed, clumps are easy to resuspend and population doubling speed and shape of cells is the same there is chance that you will see no differences between morphology of these cells, they simply like to stick together.
If there are differences in shape of cells under microscope and clumps are hard or impossible to resuspend i would try immunocytochemistry first, fix, perneabilize cells, stain cytosceleton with fluorescent dye and make photos. You will get much more information than from gene expression levels. You can quantify amouth of cytosceleton proteins in cells and find out it it is increased or not.....
About the genes connected with cell morphology I am not able to help.
Hope this helps a little and good luck
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