Preprint: Please note that this article has not completed peer review.

Mouse intestinal organoid time-course experiments from single cells

Denise Serra, Urs Mayr, Andrea Boni, Ilya Lukonin, Markus Rempfler, Ludivine Challet Meylan, Michael B. Stadler, Petr Strnad, Panagiotis Papasaikas, Dario Vischi, Annick Waldt, Guglielmo Roma, Prisca Liberali
DOI: 10.21203/rs.2.9220/v1

Abstract

Organoids recapitulate the self-organizing capacity of stem cells and the tissue organization of the original organ in a controlled and trackable environment. Intestinal organoids, in particular, can develop from a single cell into a fully-grown structure that contains most of the cell types, patterns, and morphogenetic properties of the adult intestine. Here we present a protocol for high-throughput organoid culture in multi-well plate format combined with high content immunofluorescence imaging and RNA extraction. Our protocol allows recording and analysis of thousands of organoids during several days of development. 


Keywords
organoid, self-organisation, high throughput microscopy, multiplexed immunofluorescence imaging, RNA extraction

Introduction

Reagents

Equipment

Procedure

Troubleshooting

Time Taken

Anticipated Results

References

Acknowledgements

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Preprint: Please note that this article has not completed peer review.

Mouse intestinal organoid time-course experiments from single cells

Denise Serra, Urs Mayr, Andrea Boni, Ilya Lukonin, Markus Rempfler, Ludivine Challet Meylan, Michael B. Stadler, Petr Strnad, Panagiotis Papasaikas, Dario Vischi, Annick Waldt, Guglielmo Roma, Prisca Liberali

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Abstract

Organoids recapitulate the self-organizing capacity of stem cells and the tissue organization of the original organ in a controlled and trackable environment. Intestinal organoids, in particular, can develop from a single cell into a fully-grown structure that contains most of the cell types, patterns, and morphogenetic properties of the adult intestine. Here we present a protocol for high-throughput organoid culture in multi-well plate format combined with high content immunofluorescence imaging and RNA extraction. Our protocol allows recording and analysis of thousands of organoids during several days of development. 


Introduction

Reagents

Equipment

Procedure

Troubleshooting

Time Taken

Anticipated Results

References

Acknowledgements

Learn more about our company.