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

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Programmed Morphogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2258))

Abstract

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.

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Abbreviations

2D:

Two dimensional

3D:

Three dimensional

AFM:

Atomic force microscopy

APTMS:

(3-Aminopropyl) trimethoxysilane

CAFs:

Cancer-associated fibroblasts

DCDMS:

Dichlorodimethylsilane

ECM:

Extracellular matrix

PDMS:

Polydimethylsiloxane

TAM:

Tumor-associated macrophages

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Acknowledgments

The authors would like to acknowledge National Science Foundation (NSF) Award Number 1914680 for supporting this study.

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Authors and Affiliations

  1. Harrington Department of Bioengineering, School of Biological and Health Systems Engineering (SBHSE), Arizona State University, Tempe, AZ, USA

    Harpinder Saini & Mehdi Nikkhah

  2. Center for Personalized Diagnostics (CPD), Biodesign Institute, Arizona State University, Tempe, AZ, USA

    Mehdi Nikkhah

Authors
  1. Harpinder Saini
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  2. Mehdi Nikkhah
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Corresponding author

Correspondence to Mehdi Nikkhah .

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Editors and Affiliations

  1. Department of Pathology, Division of Experimental Pathology, School of Medicine, University of Pittsburgh, Pittsburgh Liver Research Center, University of Pittsburgh, Department of Bioengineering Swanson School of Engineering, University of Pittsburgh, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Mo R. Ebrahimkhani

  2. Department of Bioengineering Swanson School of Engineering, Pittsburgh Liver Research Center, Department of Pathology, Division of Experimental Pathology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Joshua Hislop

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Saini, H., Nikkhah, M. (2021). Fabrication Method of a High-Density Co-Culture Tumor–Stroma Platform to Study Cancer Progression. In: Ebrahimkhani, M.R., Hislop, J. (eds) Programmed Morphogenesis. Methods in Molecular Biology, vol 2258. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1174-6_16

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  • DOI: https://doi.org/10.1007/978-1-0716-1174-6_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1173-9

  • Online ISBN: 978-1-0716-1174-6

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