0046 - Organoids as models to study probiotics

Research output: Contribution to conferenceAbstractAcademic

Abstract

For decades, scientists have exploited cancer cell lines as models to study host-pathogen interactions and intestinal functions in vitro. Such monotypic cell models have led to important discoveries but have notable limitations. Immortalized cell lines display biological variations such as aneuploidy, chromosome rearrangements or mutations leading to poorly reproducible results, even for the same cell line.
Organoids are gaining considerable interest as alternative models of the intestine due to their close resemblance to structural, cellular and functional complexity found in vivo. However, the three-dimensional geometry of stem-cell derived intestinal organoids limits easy access to the apical epithelium for investigating the influence of probiotics, bioactive and toxic compounds on barrier function and permeability. Here we present a new robust method for generating confluent intestinal cell monolayers from single-cell suspensions of enzymatically-dissociated organoids. Confluent polarised monolayers containing tight-junctions were formed in three days and could be used in experiments for up to two weeks. Multilineage differentiation of the ileal stem cells was demonstrated by immunohistochemistry, and RT-qPCR of cell-specific transcripts. Furthermore, we showed that adult stem-cell derived ileal organoids maintain location-specific transcriptional programs during long-term in vitro culture.
Original languageEnglish
Pages57
Number of pages1
Publication statusPublished - 19 Jun 2019
EventIPC2019 : International scientific conference on probiotics, prebiotics, gut microbiota, and health - Prague congress centre, Prague, Czech Republic
Duration: 17 Jun 201920 Jun 2019
Conference number: 13
https://www.probiotic-conference.net/

Conference

ConferenceIPC2019
CountryCzech Republic
CityPrague
Period17/06/1920/06/19
Internet address

Fingerprint

Organoids
Probiotics
Cell Line
Stem Cells
Host-Pathogen Interactions
Adult Stem Cells
Tight Junctions
Poisons
Aneuploidy
Intestines
Permeability
Suspensions
Epithelium
Chromosomes
Immunohistochemistry
Mutation
Neoplasms
In Vitro Techniques

Keywords

    Cite this

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    title = "0046 - Organoids as models to study probiotics",
    abstract = "For decades, scientists have exploited cancer cell lines as models to study host-pathogen interactions and intestinal functions in vitro. Such monotypic cell models have led to important discoveries but have notable limitations. Immortalized cell lines display biological variations such as aneuploidy, chromosome rearrangements or mutations leading to poorly reproducible results, even for the same cell line.Organoids are gaining considerable interest as alternative models of the intestine due to their close resemblance to structural, cellular and functional complexity found in vivo. However, the three-dimensional geometry of stem-cell derived intestinal organoids limits easy access to the apical epithelium for investigating the influence of probiotics, bioactive and toxic compounds on barrier function and permeability. Here we present a new robust method for generating confluent intestinal cell monolayers from single-cell suspensions of enzymatically-dissociated organoids. Confluent polarised monolayers containing tight-junctions were formed in three days and could be used in experiments for up to two weeks. Multilineage differentiation of the ileal stem cells was demonstrated by immunohistochemistry, and RT-qPCR of cell-specific transcripts. Furthermore, we showed that adult stem-cell derived ileal organoids maintain location-specific transcriptional programs during long-term in vitro culture.",
    keywords = "permeability, intestinal models, organoids, stem-cells, tight-junctions",
    author = "{van der Hee}, B. and L.M.P. Loonen and N. Taverne and J.J. Taverne-Thiele and H. Smidt and J.M. Wells",
    year = "2019",
    month = "6",
    day = "19",
    language = "English",
    pages = "57",
    note = "IPC2019 : International scientific conference on probiotics, prebiotics, gut microbiota, and health ; Conference date: 17-06-2019 Through 20-06-2019",
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    0046 - Organoids as models to study probiotics. / van der Hee, B.; Loonen, L.M.P.; Taverne, N.; Taverne-Thiele, J.J.; Smidt, H.; Wells, J.M.

    2019. 57 Abstract from IPC2019 , Prague, Czech Republic.

    Research output: Contribution to conferenceAbstractAcademic

    TY - CONF

    T1 - 0046 - Organoids as models to study probiotics

    AU - van der Hee, B.

    AU - Loonen, L.M.P.

    AU - Taverne, N.

    AU - Taverne-Thiele, J.J.

    AU - Smidt, H.

    AU - Wells, J.M.

    PY - 2019/6/19

    Y1 - 2019/6/19

    N2 - For decades, scientists have exploited cancer cell lines as models to study host-pathogen interactions and intestinal functions in vitro. Such monotypic cell models have led to important discoveries but have notable limitations. Immortalized cell lines display biological variations such as aneuploidy, chromosome rearrangements or mutations leading to poorly reproducible results, even for the same cell line.Organoids are gaining considerable interest as alternative models of the intestine due to their close resemblance to structural, cellular and functional complexity found in vivo. However, the three-dimensional geometry of stem-cell derived intestinal organoids limits easy access to the apical epithelium for investigating the influence of probiotics, bioactive and toxic compounds on barrier function and permeability. Here we present a new robust method for generating confluent intestinal cell monolayers from single-cell suspensions of enzymatically-dissociated organoids. Confluent polarised monolayers containing tight-junctions were formed in three days and could be used in experiments for up to two weeks. Multilineage differentiation of the ileal stem cells was demonstrated by immunohistochemistry, and RT-qPCR of cell-specific transcripts. Furthermore, we showed that adult stem-cell derived ileal organoids maintain location-specific transcriptional programs during long-term in vitro culture.

    AB - For decades, scientists have exploited cancer cell lines as models to study host-pathogen interactions and intestinal functions in vitro. Such monotypic cell models have led to important discoveries but have notable limitations. Immortalized cell lines display biological variations such as aneuploidy, chromosome rearrangements or mutations leading to poorly reproducible results, even for the same cell line.Organoids are gaining considerable interest as alternative models of the intestine due to their close resemblance to structural, cellular and functional complexity found in vivo. However, the three-dimensional geometry of stem-cell derived intestinal organoids limits easy access to the apical epithelium for investigating the influence of probiotics, bioactive and toxic compounds on barrier function and permeability. Here we present a new robust method for generating confluent intestinal cell monolayers from single-cell suspensions of enzymatically-dissociated organoids. Confluent polarised monolayers containing tight-junctions were formed in three days and could be used in experiments for up to two weeks. Multilineage differentiation of the ileal stem cells was demonstrated by immunohistochemistry, and RT-qPCR of cell-specific transcripts. Furthermore, we showed that adult stem-cell derived ileal organoids maintain location-specific transcriptional programs during long-term in vitro culture.

    KW - permeability

    KW - intestinal models

    KW - organoids

    KW - stem-cells

    KW - tight-junctions

    M3 - Abstract

    SP - 57

    ER -