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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
Denise Serra
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
ORCiD: https://orcid.org/0000-0003-0332-8127
Urs Mayr
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
ORCiD: https://orcid.org/0000-0001-9842-5776
Andrea Boni
Viventis Microscopy Sàrl, EPFL Innovation Park, Lausanne, Switzerland
Ilya Lukonin
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Markus Rempfler
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Ludivine Challet Meylan
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Michael B. Stadler
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Petr Strnad
Viventis Microscopy Sàrl, EPFL Innovation Park, Lausanne, Switzerland
Panagiotis Papasaikas
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Dario Vischi
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Annick Waldt
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
Guglielmo Roma
Novartis Institutes for BioMedical Research, Novartis Pharma AG, Basel, Switzerland
Prisca Liberali
Friedrich Miescher Institute for Biomedical Research (FMI), Basel, Switzerland
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
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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.
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