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In Vitro Miniaturized Tuberculosis Spheroid Model.

Shilpaa Mukundan1, Pooja Singh2, Aditi Shah1

  • 1Department of Biomedical Engineering, Rutgers, The State University of New Jersey, Jersey City, NJ 08854, USA.

Biomedicines
|September 28, 2021
PubMed
Summary
This summary is machine-generated.

We developed novel tuberculosis (TB) spheroids using human cells that mimic disease pathology, including cell death and hypoxia. This new model aids in understanding TB and discovering new drug therapies.

Keywords:
MALDI MSIin vitro modellipid characterizationtuberculosis

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Area of Science:

  • Biomedical Engineering
  • Infectious Disease Research
  • Cell Biology

Background:

  • Tuberculosis (TB) affects millions globally, posing a significant public health challenge.
  • Existing in vitro models for TB lack key pathological features like caseation, limiting their utility in drug development.
  • Effective drug therapies are hampered by the limitations of current experimental models.

Purpose of the Study:

  • To create a novel, high-throughput in vitro model for studying tuberculosis (TB).
  • To develop a model that replicates key pathological features of TB, including caseation and hypoxia.
  • To establish a platform for improved therapeutic discovery and disease modeling in TB.

Main Methods:

  • Generation of THP-1 human monocyte/macrophage spheroids infected with Mycobacterium tuberculosis (Mtb).
  • Analysis of spheroid characteristics including cell death, hypoxia, pro-inflammatory factors (TNFα), growth factors (G-CSF, VEGF), and lipid deposition via MALDI mass spectrometry imaging.
  • Adaptation of the spheroid model to include peripheral blood mononuclear cells (PBMCs) and lung fibroblasts (NHLF) in 3D co-cultures.

Main Results:

  • TB spheroids successfully recapitulated a central core of dead cells with co-localized mycobacteria and hypoxia.
  • Infected spheroids showed elevated levels of TNFα, G-CSF, and VEGF compared to controls.
  • Lipid deposition was significantly observed in TB spheroids.
  • Differential virulence strains of Mtb exhibited varied responses to Isoniazid and Rifampicin within the spheroid model.

Conclusions:

  • The developed TB spheroid model accurately mimics critical aspects of human TB pathology.
  • This model offers a promising platform for advancing TB research and accelerating the discovery of new therapeutic strategies.
  • The adaptability of the spheroid model to co-cultures suggests broad applicability in infectious disease modeling.