Initiation, Expansion, and Cryopreservation of Human Primary Tissue-Derived Normal and Diseased Organoids in Embedded Three-Dimensional Culture

Curr Protoc Cell Biol. 2019 Mar;82(1):e66. doi: 10.1002/cpcb.66. Epub 2018 Sep 28.


Organoids are primary patient-derived micro tissues grown within a three-dimensional extracellular matrix that better represents in vivo physiology and genetic diversity than existing two-dimensional cell lines. Organoids rely on the self-renewal and differentiation of tissue-resident stem cells that expand in culture and self-organize into complex three-dimensional structures. Depending on the tissue, organoids typically lack stromal, vascular, neural, and immune cells but otherwise can contain cells from all the respective tissue-specific cell lineages found in vivo. Established organoids can be initiated from cryopreserved material, cultured using largely traditional cell culture techniques and equipment, and then expanded and cryopreserved for future use. Organoid models have been developed from a variety of diseased and normal tissues including small intestine, colon, mammary, esophagus, lung, prostate, and pancreas. © 2018 by John Wiley & Sons, Inc.

Keywords: 3D; cancer; disease model; extracellular matrix; microenvironment; organoid; primary cells; stem cells; three-dimensional.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cryopreservation*
  • Extracellular Matrix / chemistry
  • Humans
  • Organoids / cytology*
  • Organoids / pathology*