3 replies to this topic

[swizla21]

HI,

Ive just started researching into THP cells, more specifically their chemotaxis towards chemoattractants using the transwell membrane plates once the THP have been activated. There are many ways to activate the THP cells.

Does anyone have any suggestions as to the best method to activate them into the macrophage phenotype?

[AntonAleraLabs]

Look into using 10 nM retinoic acid to turn THP-1 into macrophages. You need to treat THP-1 cells for 72h in 10 nM retinoic acid to turn them into macrophages. Large proportion of cells will die if you protocol is working. For activation, LPS is second to none.

Anton
Principal Consultant, Molecular and Cell Biology

Phone: (518) 334-0922
E-mail: antonb@aleralabs.com

Web: www.aleralabs.com

Alera Labs
P.O. Box 13019
Research Triangle Park, NC 27709

[victorius]

Phorbol ester, PMA, is commonly used to turn THP-1 cells into macrophage-like cells (see paper below). Also, vitamin D3 (100-200 nM x 3 days) can be used and will result in CD14 upregulation which is more pronounced in adherent, rather than floating, cells.

V.


Cellular and Molecular Life Sciences
Volume 47, Number 1, 22-31,
J. Auwerx
The human leukemia cell line, THP-1: A multifacetted model for the study of monocyte-macrophage differentiation

[swizla21]

as to retanoic acid and LPS what would you use to dilute them to get the concentrations you want to work with?

Thanks for your help so far! the PMA is working fine, but having trouble with my NBT assay as to getting the formation of the formazan particles!