U937 Cells
U937 Cells - Related Videos
06:53
Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation
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Cited by 4 •
2019
Here, we present a protocol to isolate the plasma membrane, cytoplasm and mitochondria of U937 cells without the use of high-speed centrifugation. This technique can be used to purify subcellular fractions for subsequent examination of protein localization via immunoblotting.
10:19
Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification
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Cited by 1 •
2021
This fractionation protocol will allow researchers to isolate cytoplasmic, nuclear, mitochondrial, and membrane proteins from mammalian cells. The latter two subcellular fractions are further purified via isopycnic density gradient.
08:15
Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System
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2025
As CRISPR-related protocols become increasingly useful and accessible, complications and obstacles can still arise under specific experimental conditions. This protocol outlines the creation of a receptor-interacting serine/threonine-protein kinase 1 (RIPK1/RIP1) knockout human cell line using CRISPR/Cas9 and highlights potential challenges encountered during this process.
09:43
Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells
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Cited by 3 •
2021
Described here is an established method to determine the extent of HIV-1 restriction by the cellular inhibitory protein SAMHD1. Human myeloid lineage U937 cells are transduced with a SAMHD1 expression vector co-expressing YFP, differentiated and then challenged with HIV-RFP. The level of restriction is determined by flow cytometry analysis.
03:20
XTT Assay: A Colorimetric Assay to Assess Cell Viability and Mitochondrial Activity
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2023
This article describes the protocol for XTT assay which is a colorimetric assay to assess cell viability and mitochondrial activity.
FAQs
PMA activates protein kinase C signaling, which promotes the transition of U937 cells toward an adherent, macrophage-like state. During this induction, cells undergo coordinated changes in morphology, adhesion, and gene expression. This makes PMA-treated cultures useful for examining how signaling activity is associated with myeloid differentiation in vitro.
The main indicators are a shift from suspension growth to adhesion, together with altered cell morphology and gene expression. These changes provide complementary evidence that the culture has acquired a macrophage-like phenotype rather than reflecting only a change in cell attachment. Researchers can therefore assess differentiation through both visible and molecular outcomes.
U937 cells allow investigators to study related myeloid states within a reproducible in vitro system. Maintaining the cells in suspension supports one experimental state, while PMA treatment promotes an adherent, macrophage-like phenotype. This controllability helps researchers examine how immune-cell behavior changes during differentiation and evaluate compounds that modify myeloid-cell activity.
A basic workflow begins by maintaining U937 cells as suspension-growing cultures. Researchers then apply PMA to induce differentiation and monitor the resulting changes in adhesion, morphology, and gene expression. The differentiated cultures can subsequently be used to investigate processes such as inflammation, phagocytosis, oxidative responses, or host-pathogen interactions.
These cultures support investigations of monocyte and macrophage biology, including immune-cell differentiation, inflammation, phagocytosis, and oxidative responses. They are also used to examine host-pathogen interactions and to evaluate compounds that modulate myeloid-cell activity. The model is especially useful when researchers need an experimentally controllable system for studying innate immune mechanisms.
Following PMA treatment, researchers can examine changes in cell attachment and morphology as observable outcomes, alongside altered gene expression as a molecular outcome. These readouts help characterize the macrophage-like state and connect phenotypic changes with myeloid biology. The same differentiated cultures can then support studies of inflammatory, phagocytic, oxidative, or host-pathogen responses.