The Monocytic Cell Line THP-1 as a Validated and Robust Surrogate Model for Human Dendritic Cells

Abstract

We have implemented an improved, cost-effective, and highly reproducible protocol for a simple and rapid differentiation of the human leukemia monocytic cell line THP-1 into surrogates for immature dendritic cells (iDCs) or mature dendritic cells (mDCs). The successful differentiation of THP-1 cells into iDCs was determined by high numbers of cells expressing the DC activation markers CD54 (88%) and CD86 (61%), and the absence of the maturation marker CD83. The THP-1-derived mDCs are characterized by high numbers of cells expressing CD54 (99%), CD86 (73%), and the phagocytosis marker CD11b (49%) and, in contrast to THP-1-derived iDCs, CD83 (35%) and the migration marker CXCR4 (70%). Treatment of iDCs with sensitizers, such as NiSO4 and DNCB, led to high expression of CD54 (97%/98%; GMFI, 3.0/3.2-fold induction) and CD86 (64%/96%; GMFI, 4.3/3.2-fold induction) compared to undifferentiated sensitizer-treated THP-1 (CD54, 98%/98%; CD86, 55%/96%). Thus, our iDCs are highly suitable for toxicological studies identifying potential sensitizing or inflammatory compounds. Furthermore, the expression of CD11b, CD83, and CXCR4 on our iDC and mDC surrogates could allow studies investigating the molecular mechanisms of dendritic cell maturation, phagocytosis, migration, and their use as therapeutic targets in various disorders, such as sensitization, inflammation, and cancer.

A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

“…When compared to original THP-1 cells our recent study confirmed the expected pronounced ability of THP-1-derived iDCs to phagocytose pathogen membrane particles such as zymosan. Furthermore, iDCs, but not undifferentiated THP-1 cells, are able to induce T cell activation via upregulation of IL-12p40 upon sensitizer treatment (41). Thus, a logical subsequent step is to prove whether the iDCs might be suitable surrogates for dermal dendritic cells.…”

“…Immature dendritic cells were generated according to the protocol described by Hölken and Teusch (41). In total, 1 x 10 6 THP-1 cells were seeded in 5 mL RPMI-1640 (Gibco, #22400089) containing 10% FBS (Gibco, #10270-106), 50 U/mL Pen-Strep (Gibco, #15140122) and 50 µM 2-mercaptoethanol (Gibco, #21985023) into a T25 flask.…”

“…Our previously published results showed that the human monocytic cell line THP-1 can be differentiated into immature dendritic cells (iDCs), displaying a sufficient ability and sensitivity to robustly identify classified proficiency chemicals and model sensitizers such as DNCB and NiSO 4 in vitro. Furthermore, as expected from antigen presenting cells, THP-1-originated iDCs have proven to phagocytose membrane components derived from pathogens such as zymosan and, finally, iDCs might be able to induce T cell activation via upregulation of IL-12p40 upon sensitizer treatment (41). Since THP-1-derived iDCs fulfill all required in vitro criteria, we aim to demonstrate in this study the functionality of stably integrated iDCs as dermal dendritic cell surrogates within a human full-thickness skin model and their subsequent molecular characterization before and after sensitizer treatment.…”

Evaluation and Mechanistic Investigation of Human Milk Oligosaccharide against SARS-CoV-2

“…THP-1 cells were cultured and maintained in RPMI 1640 GlutaMAX medium (Gibco) supplemented with 10% (v/v) heat-inactivated FBS and a 1% (v/v) penicillin and streptomycin mixture (Gibco). THP-1 cells were induced to differentiate into macrophages using 100 ng/mL Phorbol 12-myristate 13-acetate (PMA) as described previously, and 100 ng/mL recombinant human interleukin 4 (IL-4) and 100 ng/mL recombinant human granulocyte macrophage colony-stimulating factor (GM-CSF) were used to induce THP-1 cells to differentiate into dendritic cells (moDC) . All cells were cultured at 37 °C and 5% CO 2 .…”

The Toxoplasma secreted effector TgWIP modulates dendritic cell motility by activating host tyrosine phosphatases Shp1 and Shp2

“…Fresh cytokinesupplemented medium was exchanged every 2 days. On day 7, the semi-adherent and cells in suspension were harvested, consisting of immature DCs (modi ed from Holken et al [14]).…”