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Macrophage Enhancement of Tumor Cell Extravasation and Intravasation In Vitro

Alessandro Genna1, Francisco W Puerta Martinez2, John C McAuliffe3,4,5,6,7

  • 1Department of Immunology and Regenerative Biology, Weizmann Institute of Science, Rehovot, Israel.

Methods in Molecular Biology (Clifton, N.J.)
|January 1, 2026
PubMed

Insights

This study introduces a 3D in vitro model to investigate how macrophages enhance tumor cell intravasation and extravasation, crucial steps in cancer metastasis. The assay provides high-resolution insights into metastatic dissemination using breast and pancreatic cancer cell lines.

Area of Science:

  • Oncology
  • Cell Biology
  • Cancer Research

Background:

  • Macrophages are key regulators of the tumor microenvironment.
  • They promote tumor cell invasion, intravasation, and extravasation, driving metastatic progression.
  • Understanding these processes requires advanced in vitro models.

Purpose of the Study:

  • To describe a novel in vitro protocol for examining macrophage-enhanced tumor cell extravasation and intravasation.
  • To utilize a 3D stratified co-culture transendothelial migration (TEM) transwell system.
  • To demonstrate the assay's utility with breast and pancreatic tumor cell lines.

Main Methods:

  • Development of a 3D co-culture transwell system.
  • Stratified culture of macrophages and tumor cells.
  • Assessment of tumor cell transendothelial migration (intravasation and extravasation).

Main Results:

  • The developed 3D TEM assay successfully models macrophage-enhanced tumor cell extravasation and intravasation.
  • The protocol allows for high-resolution examination of metastatic dissemination.
  • The assay demonstrated utility with both breast and pancreatic cancer cell lines.

Conclusions:

  • This 3D in vitro TEM system is a valuable tool for studying macrophage roles in cancer metastasis.
  • The assay can be broadly applied to investigate various aspects of metastatic dissemination.
  • It offers a high-resolution platform for preclinical research in oncology.
Keywords:
MacrophagesMicroscopyTransendothelial migration (TEM)Tumor cell

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