monocytes (U 937) differentiation to macrophages - how can I confirm the differentiation? (Oct/16/2005 )

Hi!
I wonder if someone could answer me to some quetions concerning monocyte (U 937)to macrophage differentiation.

1)Do the differentiated cells continue to grow in number after replacing the medium with PMA with fresh medium without PMA? and what should I expect to see concernig morphology (shape and size? do they keep their round shape?)

2)Whta is the best way to confirm the differentiation of U937 to macrophages? (immunohistochemisty? or something else?)

Thanks in advance

Hello:

Your cells will continue differentiation after placing fresh medium. You should expect changes in morphology, they will loss round shape and will show spindle-like appearance. Be sure to feed your cells every 3-4 days.

Good luck!