Facilitating THP-1 macrophage studies by differentiating and investigating cell functions in polystyrene test tubes

Journal of Immunological Methods
Annabelle SmallAntonio Ferrante

Abstract

Macrophage cell lines are a useful model to explore the properties of primary macrophages. However, a major limitation in the use of these cells is that when they are differentiated, they become adherent and hence present with the same limitation as natural macrophages. The cells need to be detached and are often subjected to detachment techniques such as detachment buffers containing proteolytic enzymes or scraping with a rubber 'policeman'. These steps are time-consuming, reduce cell yields as well as cell viability and function. We have therefore investigated the possibility of differentiating the human macrophage THP-1 cell line in polystyrene FACS tubes to enable cells to be directly used for investigations by flow cytometry. Here we demonstrate that when the human macrophage cell line THP-1 are cultured in FACS tubes with phorbol myristate acetate added, they undergo differentiation into macrophages, assessed morphologically and by autofluorescence expression, in a similar manner to those cultured in tissue culture dishes. The cells can be readily washed and adjusted in concentration by centrifugation in the same tubes and can be directly tested for expression of cell surface markers and function by flow cytometry. This a...Continue Reading

Citations

Jan 13, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yomaira L UscáteguiManuel F Valero
Jan 2, 2020·ACS Central Science·Wuliang ZhangChad A Mirkin

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