I am looking to study opsonophagocytosis using THP-1 cell line from ATCC. Does the cell line require differentiation before the phagocytosis assay?
Hello Somnath Chakravorty
Not necessary. Please refer to the research article below under the sub-heading
Phagocytosis using undifferentiated THP-1 cells.
Article Opsonic phagocytosis of Plasmodium falciparum merozoites: Me...
Best.
THP-1 is a human monocytic cell line. When stimulated/treated, will gets differentiated into macrophages. Now, it depends on whether you would like to study opsonophagocytosis with either monocytes or macrophages. If your goal is to study opsonophagocytosis using macrophages, then THP-1 cells need to be differentiated !
BW
THP1 is a monocytic cell line and monocytes usually have low to minimal phagocytic capacity. You may differentiate monocytes to macrophages using 100-150 nM PMA and investigate phagocytosis. Please let me know if you need further help.
Bhupesh Singla Thank you for the reply, do you have a protocol that you can share with me for the differentiation and downstream harvest till start of the phagocytosis? It would be greatly appreciated.
Be careful regarding the use of THP-1 as a monocyte model. Be sensitive to the origin to the cell line THP-1 that was derived from the peripheral blood of a childhood case of acute monocytic leukemia (M5 subtype) and thus is a leukemic cell line with monocytic markers (1). As compared to primary monocytes, THP-1 cells express low levels of CD14 (2) and as such may be a poor model for primary monocytes responding to the classic activating agent LPS (3). Other differences have been reported between THP-1 and PBMCs including migration in response to chemoattractants in a transwell assay, with varying sensitivity to MCP-1, MIP-1α and LTB-4. In summary, THP-1 cells are not a monocyte cell line.
1) Tsuchiya S, Yamabe M, Yamaguchi Y, et al. Establishment and characterization of a human acute monocytic leukemia cell line (THP-1). Int J Cancer1980;26:171-6
2) Bosshart H, Heinzelmann M. Lipopolysaccharide-mediated cell activation without rapid mobilization of cytosolic free calcium. Mol Immunol2004;41:1023-8.
3) THP-1 cells as a model for human monocytes. Bosshart H, et al. Ann Transl Med. 2016
4) Comparative genotypic and phenotypic analysis of human peripheral blood monocytes and surrogate monocyte-like cell lines commonly used in metabolic disease research. Riddy DM, et al. PLoS One. 2018