The Organoid Culture Handbook

The Organoid Culture Handbook

An organoid is a miniaturized version of an organ produced in vitro that shows realistic micro-anatomy, is capable of self-renewal and self-organization, and exhibits similar functionality as the tissue of origin.

Organoids can be generated from adult tissue-derived stem cells (ASCs) or from embryonic (ESCs) and induced pluripotent stem cells (iPSCs). Organoid culture offers advancements over classical 3D systems, such as spheroids, because they can develop into complex multi-cellular tissues with self-organized compartmentalized regions, including the potential for vasculature and immune cells.

Researchers have devised methods to generate physiologically relevant organoid models for many organs, including the intestines, lung, brain, liver, lung, pancreas, and heart. While techniques and protocols for generating, processing, and imaging organoids are still evolving, they are being accepted as the standard models for interrogating basic organ biology, disease modeling, and tissue regeneration.

The Organoid Handbook provides a tissue-specific overview of:

  • Landmark publications highlighting organoid technology
  • Key reagents for organoid and 3D cell culture
  • Protocols for organoid models
  • Troubleshooting organoid cultures
  • And more!

The Organoid Culture Handbook

Preview of What's to Learn in This Informational Guide

Brain Organoids

Protocols to generate 3D brain organoids from ESCs and iPSCs were first published in 2009. These studies showed that pluripotent stem cells could differentiate into cerebral organoids containing specific cortical regions, neural progenitor populations, and cortical layer patterning. Cerebral organoids have since been employed to uncover evolutionary differences in brain development between species, mechanisms of brain region interconnectivity, and the developmental physiology of normal and diseased brain regions. iPSC-derived organoids show great potential for use in drug discovery as well as modeling neurodegenerative disease and viral brain infection.


Neural Progenitor Cells in Cerebral Organoids. (Right) Human iPSCs were differentiated into cerebral organoids following the Lancaster protocol. Organoids were harvested and stained using a Human Pax6 Polyclonal Antibody (red; Catalog # AF8150) to identify neural progenitor cells in the developing cerebral tissue. Tissue was counterstained using DAPI (blue; Catalog # 5748).

Optimizing Organoid Culture Conditions

Notable Publications and Protocols

Publication Description Bio-Techne Products Used
Pollen, A.A. et al. (2019) Cell 176:743. Establishing Cerebral Organoids as Models of Human-Specific Brain Evolution Y-27632 dihydrochloride, Cat# 1254, Selective ROCK inhibitor

SB 431542, Cat# 1614, Selective TGF-βRI inhibitor
Bershteyn, M. et al. (2017) Cell Stem Cell 20:435. Human iPSC-Derived Cerebral Organoids Model Cellular Features of Lissencephaly and Reveal Prolonged Mitosis of Outer Radial Glia Y-27632 dihydrochloride, Cat# 1254, Selective ROCK inhibitor

SB 431542, Cat# 1614, Selective TGF-βRI inhibitor
Bagley, J. A. et al. (2017) Nature Methods 13:743. Fused cerebral organoids model interactions between brain regions  
Lancaster M.A. and J. A. Knoblich (2014) Nat. Protocols. 9:2329. Generation of cerebral organoids from human pluripotent stem cells  

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