Fabrication Method of a High-Density Co-Culture Tumor–Stroma Platform to Study Cancer Progression

Harpinder Saini, Mehdi Nikkhah

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations


Cancer has now been established as one of the most common chronic diseases due to high mortality rate. The early stage of non-invasive tumors can now be successfully treated leading to have high survival rates; however, the late stage invasive and metastatic tumors still suffer from poor treatment outcomes. Among multiple contributing factors, the role of tumor microenvironment and its complexities has been well recognized in cancer progression. Stromal cells including cancer-associated fibroblasts (CAFs), endothelial cells, adipocytes, immune cells as well as extracellular matrix (ECM) continuously interact with malignant cells and regulate various hallmarks of cancer including tumor growth, invasion, and intravasation. To better understand the role of the interaction between tumor cells and their surrounding microenvironment, numerous model systems ranging from two-dimensional (2D) assays to 3D hydrogels and in vivo murine xenografts have been utilized. While each one of these model systems exhibit certain advantages in studying biological facets of tumor progression, they are often limited to perform well-controlled mechanistic studies due to various factors including lack of tumor–stroma organotypic organization and presence of confounding biochemical and biophysical factors within the tumor microenvironment. In this regard, in the past few years, 3D in vitro microengineered model systems are becoming instrumental to precisely mimic the complexities of the native tumor microenvironment to conduct fundamental and well-designed studies for multiple purposes ranging from biological discovery to therapeutic screening. These model systems include microfluidics, micro-patterned features, and 3D organoids. In this chapter, we will outline the fabrication strategy of our microengineered 3D co-culture tumor–stromal model which comprises high-density array of tumor seeded microwells surrounded by stromal cells, such as CAFs encapsulated within collagen-based hydrogel. The developed platform provides excellent spatial organization of tumor and stromal entities with designated initial architecture and cellular positioning, therefore enabling to study the specific role of cell–cell and cell–ECM interaction on tumor proliferation/expansion, cancer cell migration as well as stromal activation. The developed platform is compatible with standard biological assays enabling gene and protein expression analyses across different types of cancer and co-culture of tumor and stromal cells.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages15
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Co-Culture
  • Microengineered systems
  • Micromolding
  • Tumor microenvironment
  • Tumor progression
  • Tumor–stroma model

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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