Clinostat 3D Cell Culture: Protocols for the Preparation and Functional Analysis of Highly Reproducible, Large, Uniform Spheroids and Organoids

Krzysztof Wrzesinski*, Helle Sedighi Frandsen, Carlemi Calitz, Chrisna Gouws, Barbara Korzeniowska, Stephen J. Fey

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterCommunication

Abstract

Growing cells as 3D structures need not be difficult. Often, it is not necessary to change cell type, additives or growth media used. All that needs to be changed is the geometry: cells (whether primary, induced pluripotent, transformed or immortal) simply have to be grown in conditions that promote cell–cell adhesion while allowing gas, nutrient, signal, and metabolite exchange. Downstream analysis can become more complicated because many assays (like phase contrast microscopy) cannot be used, but their replacements have been in use for many years. Most importantly, there is a huge gain in value in obtaining data that is more representative of the organism in vivo. It is the goal of the protocols presented here to make the transition to a new dimension as painless as possible. Grown optimally, most biopsy derived organoids will retain patient phenotypes, while cell (both stem cell, induced or otherwise or immortalized) derived organoids or spheroids will recover tissue functionality.

Original languageEnglish
Title of host publicationNext Generation Culture Platforms for Reliable In Vitro Models : Methods and Protocols
EditorsTiziana A.L. Brevini, Alireza Fazeli, Kursad Turksen
PublisherHumana Press
Publication date2021
Pages17-62
ISBN (Print)978-1-0716-1245-3
ISBN (Electronic)978-1-0716-1246-0
DOIs
Publication statusPublished - 2021
SeriesMethods in Molecular Biology
Volume2273
ISSN1064-3745

Keywords

  • Biopsy
  • Bioreactor
  • Cell lines
  • Clinostat
  • Coculture
  • High reproducibility
  • High yield
  • Long-term culture
  • Organoids
  • Protocols
  • Spheroids
  • Stem cells
  • Cell Culture Techniques/methods
  • Humans
  • Spheroids, Cellular/cytology
  • Cell Differentiation/physiology
  • Organoids/cytology
  • Stem Cells/cytology

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