Establishment of a Patient-Derived, Magnetic Levitation-Based, Three-Dimensional Spheroid Granuloma Model for Human Tuberculosis

Abstract

TB is a highly infectious disease, with granulomas as its hallmark. Granulomas play an important role in the control of M. tuberculosis infection and as such are crucial indicators for our understanding of host resistance to TB.

Magnetic-Based Human Tissue 3D Cell Culture: A Systematic Review

“…Magnetic-based models allow us to effectively produce homogeneous 3D aggregates and with desirable compactness [ 1 , 17 ]. Of the twenty-five studies included in this review, nine studies used only magnetic levitation [ 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ], eleven studies used only the bioprinting method [ 14 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 ], and three studies used both methods [ 15 , 36 , 37 ]. Only two studies used the magnetic ring formation method [ 22 , 38 ].…”

“…The 24-well plates were the most used for studies using the levitation method; whereas, the 96-well plates were the most used for the bioprinting method. More precisely, nine studies used the 24-well Bio-Assembler Kit [ 15 , 23 , 24 , 25 , 26 , 30 , 32 , 37 , 39 ], eight studies used the 96-well Bioprinting Kit [ 14 , 27 , 28 , 29 , 31 , 33 , 35 , 36 ], two studies used both 6-well and 24-well Bio-Assembler Kit [ 17 , 19 ], one study used only the 6-well Bio-Assembler Kit [ 34 ], and one study used the 96-well Ring Drive for ring structure formation [ 38 ]. There is an additional study that used tissue culture Petri dishes covered by a top cover with an attached neodymium magnet [ 22 ].…”

“…M3D cultures of human pancreatic β cells (endoC-βH3 cell line) and VEGF pre-screened umbilical vein endothelial cells (HUVEC cell line) allowed for the investigation of the interaction between both cell types [ 31 ]. Tuberculosis granulomas were resembled into m3D cultures using human alveolar macrophages and autologous CD 3+ T cells to study the host/pathogen pathways and the immune response involved in the infection process [ 37 ]. Chicken hepatocytes and non-parenchymal cells were used to establish a proper hepatic inflammatory model for testing potential proinflammatory molecules [ 35 ].…”

“…These tools are mostly applicable for: (i) analyzing cell proliferation, viability, and metabolism; (ii) evaluating the morphology, size, volume, sphericity, and aggregation of the m3D structures; (iii) quantifying the global DNA content and the expression levels of mRNA; (iv) analyzing the intercellular communications and the formation of ECM; and (v) assessing the maintenance of phenotype and function, among others. Almost all studies included histological analysis by immunohistochemistry (IHC) and fluorescence, confocal, and electron microscopy, followed by image analysis in ImageJ software [ 13 , 15 , 17 , 19 , 23 , 24 , 26 , 27 , 29 , 30 , 31 , 32 , 37 , 39 ]. These techniques can be considered the standard techniques for analyzing all types of m3D cultures, since the MNP does not interfere with immunofluorescent assays, either increasing signal noise or interfering with fluorophore detection [ 13 ].…”

Spatial Resolution of Mycobacterium tuberculosis Bacteria and Their Surrounding Immune Environments Based on Selected Key Transcripts in Mouse Lungs

“…In vitro generated granulomas would give the opportunity to manipulate and challenge the effect of identified protective expression patterns. Although important progress in the in vitro granuloma field has been achieved, generating, e.g., three-dimensional spheroid human granulomas forming single organized structures consisting of human lung-derived alveolar macrophages surrounded by layer of autologous T cells to study innate and adaptive stages of the TB granuloma, the reduced complexity of in vivo granulomas must be considered ( 50 , 51 ). Lastly, a transcriptomic meta-analysis of peripheral blood from patients with active and latent TB could correlate identified patterns of Mtb’s immune environments during disease progression to systemic measurable parameters.…”