I will be inducing differentiation of GM colony forming units (from CFC assay) using M-CSF and IL-1beta in medium to yield promonocytes (harvest cells early before complete maturation, I know not of any other methods yet).

A comparison between these promonocytes with parental THP-1 will then be made to ascertain similarity.

Following that, I intend to further differentiate these cells into macrophages. So, what is the best method to achieve it? I am aware of having to add 100ng/ml PMA provided they are similar to THP-1 cells (thereby using the same differentiation protocol as THP-1 differentiation).

If they prove dissimilar to THP-1, would it be better to use a different differentiation approach with just M-CSF and IL-1beta without PMA? Should I allow monocytes to adhere or cultivate on poly-HEMA?

Any detailed protocols would be of much help as well.

Thanks.

Wei Xing