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Controlling Cell Organization in 3D Coculture Spheroids Using DNA Interactions.

Sven A Saemundsson1, Saheli Ganguly1, Shane D Curry1

  • 1Department of Chemical and Biological Engineering, University of Colorado, Boulder, 596 UCB, Boulder, Colorado 80303, United States.

ACS Biomaterials Science & Engineering
|May 8, 2023
PubMed
Summary

Researchers used DNA hybridization to control cell distribution in 3D tumor spheroids. This method achieved uniform mixing of cancer cells and fibroblasts, influencing extracellular matrix production and offering insights into tumor progression.

Keywords:
3D cell organizationDNA hybridizationcoculturefibronectin

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Area of Science:

  • * Oncology
  • * Biomaterials Science
  • * Cell Biology

Background:

  • * The tumor microenvironment (TME) is crucial for tumor progression and drug resistance, involving complex interactions between cancer cells and stromal/immune cells.
  • * Current in vitro 3D coculture models struggle to precisely control the spatial organization of different cell types within spheroids.
  • * Precise control over cellular arrangement is needed to accurately model TME dynamics and develop effective anticancer therapies.

Purpose of the Study:

  • * To develop a novel in vitro 3D coculture tumor model with controlled spatial organization of heterotypic cells.
  • * To investigate the impact of DNA-mediated cell-cell interactions on spheroid formation and composition.
  • * To assess how defined cellular arrangements influence extracellular matrix (ECM) production and cell-cell interactions.

Main Methods:

  • * Engineered 3D coculture tumor spheroids using MDA-MB-468 breast cancer cells and NIH/3T3 fibroblasts.
  • * Modulated cell-cell interactions via complementary DNA hybridization to direct spatial organization.
  • * Analyzed spheroid composition using microscopy and quantified ECM protein production (fibronectin, E-cadherin) via antibody staining.

Main Results:

  • * DNA hybridization successfully directed uniform mixing of cancer cells and fibroblasts within spheroids, preventing typical cell sorting.
  • * Coculture spheroids with uniform cell distribution exhibited significantly higher fibronectin secretion compared to those with segregated cell populations.
  • * E-cadherin levels remained similar across different spheroid compositions, suggesting specific influences on matrix production.

Conclusions:

  • * DNA templating offers a method to precisely control heterotypic cell distribution in 3D coculture spheroids.
  • * Controlled cellular organization within spheroids can significantly alter ECM protein production, impacting TME characteristics.
  • * This model provides a platform for mechanistic studies on how cellular arrangement influences tumor progression, metastasis, and drug resistance.