Accelerated production of human epithelial organoids in a miniaturized spinning bioreactor
Shicheng Ye, Ary Marsee, Gilles van Tienderen, Mohammad Rezaeimoghaddam, Hafsah Sheikh, Roos‐Anne Samsom, Eelco J.P. de Koning, Sabine A. Fuchs, Monique M.A. Verstegen, Luc J. W. van der Laan, Frans N. van de Vosse, Jos Malda, Keita Ito, Bart Spee, Kerstin Schneeberger
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.