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Engineering and Evaluating Vascularized Organotypic Spheroids On-Chip.

James J Tronolone1, Nadin Mohamed1, Christopher P Chaftari1

  • 1Department of Biomedical Engineering, College of Engineering, Texas A&M University, College Station, Texas.

Current Protocols
|November 21, 2024
PubMed
Summary

This study provides a standardized protocol for vascularizing organotypic spheroids, crucial for advanced in vitro models. The guide ensures reproducibility for researchers developing vascularized organoid-based microphysiological platforms.

Keywords:
microphysiological systemorganoidorgan‐chipspheroidvascularized

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Area of Science:

  • Biotechnology
  • Tissue Engineering
  • Microfluidics

Background:

  • Organotypic spheroids are valuable in vitro models but require vascularization for physiological relevance.
  • Current vascularization methods lack standardization and reproducibility, limiting their widespread adoption.
  • Developing standardized protocols is essential for advancing organoid-based research.

Purpose of the Study:

  • To provide a detailed, step-by-step guide for vascularizing organotypic spheroids.
  • To standardize protocols for microfluidic chip design, spheroid fabrication, and vascularization techniques.
  • To enhance the reproducibility and adoption of vascularized organoid models.

Main Methods:

  • Microfluidic chip design and fabrication.
  • Organotypic spheroid fabrication techniques.
  • Vascularization via vasculogenesis and angiogenesis, including molecular assays and computational modeling.

Main Results:

  • A systematic approach to vascularize organotypic spheroids (pancreatic islets and cancer spheroids) is presented.
  • Protocols cover chip design, spheroid fabrication, and on-chip vascularization.
  • Methods for characterization, quantification, and mass transport analysis are included.

Conclusions:

  • The developed protocols contribute to standardization and reproducibility in vascularized organoid research.
  • This guide aims to enhance the adoption of organoid-based microphysiological platforms by researchers.
  • Standardized vascularization is key to improving the longevity, stability, and relevance of in vitro models.