Microbiology JoVE (Open Access) Citable · DOI

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

Name: A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection
Uploaded: 2026-06-17
Duration: 10 min 10 s
Description: Tuberculosis (TB) still holds a major threat to the health of people worldwide, and there is a need for cost-efficient but reliable models to help us ...

DOI: 10.3791/53084-v

What you'll learn

✓ Construct a 3D human lung tissue model with fibroblasts and collagen matrix
✓ Seed immune and epithelial cells into the 3D lung tissue model
✓ Visualize and quantify granuloma formation and immune cell migration
✓ Analyze mycobacterial infection dynamics in physiological lung tissue

Protocol

Human tuberculosis infection is a complex process, which is difficult to model in vitro. Here we describe a novel 3D human lung tissue model that recapitulates the dynamics that occur during infection, including the migration of immune cells and early granuloma formation in a physiological environment.

Difficulty

advanced

Total time

~5-7 days (including continuous culture phase and infection observation)

Model organism

Human primary/cultured cells (fibroblasts, epithelial cells, immune cells)

Steps

Establish fibroblast-collagen matrix continuous culture

Prepare and maintain a fibroblast-embedded collagen matrix as the foundational 3D tissue scaffold. This layer provides structural support and mimics the lung extracellular matrix.

▶ 01:13

Seed immune and lung epithelial cells

Add lung epithelial cells and immune cells (macrophages, T cells) to the fibroblast-collagen matrix to create a physiologically relevant cellular composition that recapitulates lung tissue architecture.

▶ 03:35

Expose tissue to air and prepare for imaging

Culture the 3D lung tissue model at the air-liquid interface, harvest the tissue construct, and mount it on appropriate imaging platforms for microscopy analysis.

▶ 05:42

Visualize, acquire and analyze tissue quantitatively

Perform confocal or multi-photon microscopy to visualize immune cell migration and granuloma formation; conduct quantitative image analysis to measure infection dynamics and tissue responses.

▶ 07:12

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