Cell Biology JoVE (Open Access) Citable · DOI

Long-term Culture of Human Breast Cancer Specimens and Their Analysis Using Optical Projection Tomography

Name: Long-term Culture of Human Breast Cancer Specimens and Their Analysis Using Optical Projection Tomography
Uploaded: 2026-06-23
Duration: 10 min 10 s
Description: Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later ...

DOI: 10.3791/3085-v

What you'll learn

✓ Extract and prepare collagen scaffolds from rat tail tissue
✓ Establish long-term 3D breast cancer culture assays from patient specimens
✓ Apply Optical Projection Tomography to quantify tumor expansion in three dimensions
✓ Assess drug response variability across breast cancer subtypes

Protocol

We have developed a collagen-based in vitro assay which promotes proliferation and invasion from samples of all breast cancer subtypes. Optical Projection Tomography, a three dimensional microscopy technique was utilised to visualise and quantify tumour expansion. This assay may be used to quantify drug response of individual tumour samples.

Difficulty

advanced

Total time

~5–7 days per specimen (collagen extraction ~1 day, culture establishment ~3–5 days, imaging and reconstruction ~1–2 days)

Model organism

Human breast cancer specimens; Rat (collagen source)

Biosafety

BSL-2

Steps

Extract collagen from rat tail tissue

Isolate collagen fibrils from rat tails using acid extraction and precipitation methods. This collagen serves as the scaffold matrix for the 3D culture assay.

▶ 01:25

Create three-dimensional tumor culture assays

Embed human breast cancer specimens within collagen-based matrices and establish culture conditions that promote tumor cell proliferation and invasion in three dimensions.

▶ 02:51

Perform antibody staining for optical analysis

Apply fluorescent antibodies to tumor samples to mark relevant cellular structures and markers prior to three-dimensional imaging and reconstruction.

▶ 05:14

Acquire and reconstruct optical projection tomography

Use Optical Projection Tomography to acquire high-resolution three-dimensional microscopy data and reconstruct volumetric images for quantitative tumor expansion analysis.

▶ 07:07

Analyze representative three-dimensional constructions

Review and interpret reconstructed 3D tumor models to visualize spatial growth patterns and quantify drug response across different breast cancer subtypes.

▶ 09:27

💬 Comments coming soon