Dear experts,
I would like to work on THP-1 as adherent cells without changing its natural. Is there any method or protocol for develop adherence cell from suspension cell lines?
Hello Dr.Stalin! In our laboratory we promote their adhesion by addition of phorbol 12-myristate 13-acetate (PMA) at a concentration of 100 ng/ml. This conducts the differentation of THP-1 monocytes into macrophages. However, when you say "without changing its natural I do not know if you mean to preserve their monocyte phenotype". THP-1 cells are non-adherent, so all treatments you perform to promote their attachment are going to have an impact on them. For instance, PMA tends to upregulate the expression of some genes in differentiated macrophages, which could affect the gene expression induced by other stimuli. If I am not wrong and I remember well, the treatment enhances inflammatory genes through NFKB pathway. In our case, as we are studying inflammation, we added a period of arrest, leaving the cells without PMA in order to reduce that possible enhanced expression.
I would recommend you (depending on the experiments you have to perform) to try different concentrations of PMA in order to add the minimum necessary to promote their adhesion without enhancing the gene expression. Firstly, I would check the expression of inflammatory genes after the treatment and after the arrest, to see how the treatment affected your cells and whether the arrest decreased the inflammatory response. Lastly, I would characterize the cells through flow cytometry or gene expression of monocyte/macrophage markers to see the profile of your cells.
You can treat the plate or the dish where you want to seed them with Poli-L-Lysine and then let it incubate for 1 hour or even less at 37ºC 5%CO2; you can also do it in a IBIDI treated plate. At least it has worked for me in human primary white cells extracted from peripheral blood although the adherence is not the stiffest.
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