I need some suggestions about differentiation process of U-937 cell line. It would be about seeding concentration of cell, PMA concentration for treatment or resting step ...
If you have, could you send to me cell image for each step...
Hi Adem,
You can differentiate U-937 cell line using "a differentiatial mark" that we had utilized in our experiments with PMA-stimulated U937 macrophageal cell line. Namely, as U937 cells respond to the presence of LPS with a marked increase in the rate of cellular AA metabolism and cytokine release into the extracellular space, we measured activity of cyclooxygenase 2 (COX), which catalyze the key step in the conversion of cellular arachidonic acid into prostaglandins. The COX-2 expression is strictly coordinated with up-regulated in PMA-differentiated U937 cells stimulated with lipopolysaccharide (LPS). At the same time, COX-1 is not up-regulated at all. In turn, another "market sign" we'd used from time to time was enzymic activity of the Mg+2-dependent phosphatidic acid phosphohydrolase 1 (PAP-1), because up-regulation of COX-2 strictly depends on the PAP-1.
In regard to your request on seeding concentration of cell, PMA concentration for treatment, and on phenotypic and morphologic changes observed in U937 cells after treatment with PMA (100 nM) in the presence of Haemophilus influenzae, please read this paper of Dr. Jahn and colleagues
https://www.researchgate.net/profile/Andreas_Jahn1/publication/286397533/figure/fig1/AS:306499189526528@1450086509048/W640.jpg
Best wishes,
Ilya
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