Accelerated production of human epithelial organoids in a miniaturized spinning bioreactor

Cell Rep Methods. 2024 Nov 18;4(11):100903. doi: 10.1016/j.crmeth.2024.100903.

Abstract

Conventional static culture of organoids necessitates weekly manual passaging and results in nonhomogeneous exposure of organoids to nutrients, oxygen, and toxic metabolites. Here, we developed a miniaturized spinning bioreactor, RPMotion, specifically optimized for accelerated and cost-effective culture of epithelial organoids under homogeneous conditions. We established tissue-specific RPMotion settings and standard operating protocols for the expansion of human epithelial organoids derived from the liver, intestine, and pancreas. All organoid types proliferated faster in the bioreactor (5.2-fold, 3-fold, and 4-fold, respectively) compared to static culture while keeping their organ-specific phenotypes. We confirmed that the bioreactor is suitable for organoid establishment directly from biopsies and for long-term expansion of liver organoids. Furthermore, we showed that after accelerated expansion, liver organoids can be differentiated into hepatocyte-like cells in the RPMotion bioreactor. In conclusion, this miniaturized bioreactor enables work-, time-, and cost-efficient organoid culture, holding great promise for organoid-based fundamental and translational research and development.

Keywords: CP: Biotechnology; CP: Stem cell; RPMotion; bioreactor; organoid; stem cell; suspension culture.

MeSH terms

  • Bioreactors*
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cell Differentiation
  • Cell Proliferation
  • Cells, Cultured
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Hepatocytes / cytology
  • Hepatocytes / metabolism
  • Humans
  • Liver / cytology
  • Liver / metabolism
  • Miniaturization
  • Organoids* / cytology
  • Organoids* / metabolism
  • Pancreas / cytology
  • Pancreas / metabolism