A process engineering approach to increase organoid yield.

TitleA process engineering approach to increase organoid yield.
Publication TypeJournal Article
Year of Publication2017
AuthorsArora, N, Alsous, JImran, Guggenheim, JW, Mak, M, Munera, J, Wells, JM, Kamm, RD, H Asada, H, Shvartsman, SY, Griffith, LG
JournalDevelopment
Volume144
Issue6
Pagination1128-1136
Date Published2017 03 15
ISSN1477-9129
KeywordsCell Count, Cell Size, Cells, Cultured, Digestive System, Flow Cytometry, Humans, Organoids, Spheroids, Cellular, Tissue Engineering
Abstract

<p>Temporal manipulation of the environment and growth factors can direct differentiation of human pluripotent stem cells into organoids - aggregates with multiple tissue-specific cell types and three-dimensional structure mimicking native organs. A mechanistic understanding of early organoid formation is essential for improving the robustness of these methods, which is necessary prior to use in drug development and regenerative medicine. We investigated intestinal organoid emergence, focusing on measurable parameters of hindgut spheroids, the intermediate step between definitive endoderm and mature organoids. We found that 13% of spheroids were pre-organoids that matured into intestinal organoids. Spheroids varied by several structural parameters: cell number, diameter and morphology. Hypothesizing that diameter and the morphological feature of an inner mass were key parameters for spheroid maturation, we sorted spheroids using an automated micropipette aspiration and release system and monitored the cultures for organoid formation. We discovered that populations of spheroids with a diameter greater than 75 μm and an inner mass are enriched 1.5- and 3.8-fold for pre-organoids, respectively, thus providing rational guidelines towards establishing a robust protocol for high quality intestinal organoids.</p>

DOI10.1242/dev.142919
Alternate JournalDevelopment
PubMed ID28174251
PubMed Central IDPMC5358111
Grant ListR01 GM107103 / GM / NIGMS NIH HHS / United States