Engineering of three-dimensional microenvironments to promote contractile behavior in primary intestinal organoids

Integrative Biology : Quantitative Biosciences From Nano to Macro
Rebecca L DiMarcoSarah C Heilshorn

Abstract

Multiple culture techniques now exist for the long-term maintenance of neonatal primary murine intestinal organoids in vitro; however, the achievement of contractile behavior within cultured organoids has thus far been infrequent and unpredictable. Here we combine finite element simulation of oxygen transport and quantitative comparative analysis of cellular microenvironments to elucidate the critical variables that promote reproducible intestinal organoid contraction. Experimentally, oxygen distribution was manipulated by adjusting the ambient oxygen concentration along with the use of semi-permeable membranes to enhance transport. The culture microenvironment was further tailored through variation of collagen type-I matrix density, addition of exogenous R-spondin1, and specification of culture geometry. "Air-liquid interface" cultures resulted in significantly higher numbers of contractile cultures relative to traditional submerged cultures. These interface cultures were confirmed to have enhanced and more symmetric oxygen transport relative to traditional submerged cultures. While oxygen availability was found to impact in vitro contraction rate and the orientation of contractile movement, it was not a key factor in enabling...Continue Reading

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Citations

Aug 15, 2015·Advanced Functional Materials·Lei CaiSarah C Heilshorn
Sep 16, 2015·Biomaterials Science·Rebecca L DiMarcoSarah C Heilshorn
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Aug 14, 2021·Nature Materials·Bauer L LeSavageSarah C Heilshorn
Nov 3, 2020·Cell Regeneration·Yaqi LiGuoqiang Hua

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