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Am J Reprod Immunol. 2013 Jul;70(1):80-6. doi: 10.1111/aji.12129. Epub 2013 Apr 29.

Effect of culture conditions on the phenotype of THP-1 monocyte cell line.

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  • 1Department of Obstetrics Gynecology and Reproductive Science, Reproductive Immunology Unit, Yale University School of Medicine, New Haven, CT 06520, USA.

Abstract

PROBLEM:

Macrophage function has many implications in a variety of diseases. Understanding their biology becomes imperative when trying to elucidate immune cell interactions with their environment, and in vitro cell lines allow researchers to manipulate these interactions. A common cell line used is THP-1, a promyeloid cell line suggestive to outside factors, and therefore sensitive to culture conditions. In this study, we describe how culture conditions can alter THP-1 morphology and in turn affect their response to differentiation stimuli.

METHOD OF STUDY:

THP-1 cells were cultured in two conditions and treated with phorbol 12-myristate 13-acetate (PMA) or MCSF. CD14 surface expression was determined by flow cytometry and cytokine/chemokine production determined by multiplex analysis.

RESULTS:

Culture conditions of THP-1 affect their response to PMA. Highly confluent THP-1 cells differentiate into a heterogeneous population responsive to PMA as seen by an increase in CD14 expression. However, these cells, cultured in low confluence, remain as a homogenous population and do not gain CD14. Additionally, there are major differences in the constitutive cytokine profile.

CONCLUSION:

We demonstrate that the culture conditions of THP-1 cells can alter their response PMA. This suggests that culture techniques may account for the discrepancy in the literature of both basal THP-1 phenotype and their response to PMA.

© 2013 John Wiley & Sons Ltd.

PMID:
23621670
[PubMed - indexed for MEDLINE]
PMCID:
PMC3703650
Free PMC Article
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