In Vitro Miniaturized Tuberculosis Spheroid Model

Abstract

Tuberculosis (TB) is a public health concern that impacts 10 million people around the world. Current in vitro models are low throughput and/or lack caseation, which impairs drug effectiveness in humans. Here, we report the generation of THP-1 human monocyte/macrophage spheroids housing mycobacteria (TB spheroids). These TB spheroids have a central core of dead cells co-localized with mycobacteria and are hypoxic. TB spheroids exhibit higher levels of pro-inflammatory factor TNFα and growth factors G-CSF and VEGF when compared to non-infected control. TB spheroids show high levels of lipid deposition, characterized by MALDI mass spectrometry imaging. TB spheroids infected with strains of differential virulence, Mycobacterium tuberculosis (Mtb) HN878 and CDC1551 vary in response to Isoniazid and Rifampicin. Finally, we adapt the spheroid model to form peripheral blood mononuclear cells (PBMCs) and lung fibroblasts (NHLF) 3D co-cultures. These results pave the way for the development of new strategies for disease modeling and therapeutic discovery.

Harnessing graphene oxide nanocarriers for siRNA delivery in a 3D spheroid model of lung cancer

“…To the best of our knowledge, no studies have reported the formation of spheroids using NB4 cells, while THP-1 cells were observed to undergo disaggregation in a previous work, similar to our findings. 37 In this study, THP-1 cells differentiated into macrophages and formed compact spheroids, while the non-differentiated THP-1 cells were in a loose aggregate configuration after 3 d of incubation. Similar results were published with adherent cells, where NIH 3T3 formed a compact spheroid structure starting just 6 h after seeding, while A549 cells resulted in a loose aggregate during the first days of incubation.…”

In vitro modelling of bacterial pneumonia: a comparative analysis of widely applied complex cell culture models

“…Apart from endothelial cells, immune cells can be incorporated. The murine macrophage cell line J774A.1 was used in an M. pneumoniae cell coculture ALI model by Shi et al ( 2023 ), whereas human THP-1 monocytes were used in both ALI and spheroid model infected with M. tuberculosis (Reuschl et al 2017 , Mukundan et al 2021 ). Conversely, Kotze et al.…”

Initial immune response after exposure to Mycobacterium tuberculosis or to SARS-COV-2: similarities and differences

“…Therefore, a validated model that recapitulates TB in human lungs is critical to support TB research. In this regard, a number of in vitro systems ( 176 ), spheroids ( 177 ), human airway organoids ( 178 ), and experimental animal models of TB such as zebrafish ( 179 ), mouse ( 180 ), guinea pig ( 181 ), rabbit ( 182 ) and rat ( 183 ) have provided new insights into the local events that occur during few days and weeks post Mtb infection. In particular, Mtb infection in nonhuman primates closely recapitulates human TB and these models can be used to study the full spectrum of infection outcome and pathology of TB ( 184 ).…”