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IntestiCult™ Organoid Growth Medium (Human)

Cell culture medium for establishment and maintenance of human intestinal organoids

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IntestiCult™ Organoid Growth Medium (Human)

Cell culture medium for establishment and maintenance of human intestinal organoids

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Cell culture medium for establishment and maintenance of human intestinal organoids
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Product Advantages


  • Convenient, in vitro system that recapitulates many key characteristics of the adult intestinal epithelium, including intra- and intercellular signaling, self-propagating stem cell niche, and functional transport into and out of the lumen

  • Complete medium formulation that delivers consistent results

  • Enables generation of intestinal organoids in one week

  • Easy-to-use format and optimized protocol

What's Included

  • IntestiCult™ OGM Human Basal Medium, 50 mL
  • Organoid Supplement, 50 mL
    • Products for Your Protocol
    To see all required products for your protocol, please consult the Protocols and Documentation.
    1. Gentle Cell Dissociation Reagent
      Gentle Cell Dissociation Reagent

      cGMP, enzyme-free cell dissociation reagent

    2. DMEM/F-12 with 15 mM HEPES
      DMEM/F-12 with 15 mM HEPES

      Dulbecco's Modified Eagle's Medium/Nutrient Ham's Mixture F-12 (DMEM/F-12) with 15 mM HEPES buffer

    3. D-PBS (Without Ca++ and Mg++)
      D-PBS (Without Ca++ and Mg++)

      Dulbecco’s phosphate-buffered saline without calcium and magnesium

    4. Y-27632 (Dihydrochloride)
      Y-27632 (Dihydrochloride)

      RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

    What Our Scientist Says

    Organoids have truly expanded the limits of what's possible for in vitro studies of the intestinal epithelium. By providing optimized culture media and robust, approachable protocols, we are making these technologies more accessible to researchers.

    Ryan ConderAssociate Director, Gastrointestinal Biology
    Ryan Conder, Associate Director

    Overview

    Model key characteristics of the adult intestinal epithelium using intestinal organoids established and maintained with this complete medium formulation and optimized protocol. Using the easy-to-follow and robust protocol, you can derive organoids from human intestinal crypts in one week; organoid growth across donor samples, including those that are otherwise difficult to grow, is enabled by an enriched stem cell population.

    Organoids grown in IntestiCult™ Organoid Growth Medium (Human) incorporate a functional lumen enclosed by a polarized intestinal epithelial cell layer and, for versatile modeling applications, can be further differentiated in 3D or in 2D as submerged monolayers or air-liquid interface (ALI) cultures using IntestiCult™ Organoid Differentiation Medium (Human; Catalog #100-0214).

    Applications of intestinal organoid cultures include studying the development and function of intestinal epithelium, modeling intestinal diseases, and screening molecules for both efficacy and toxicity in an intestinal model. Intestinal organoid cultures can also be used for investigation of adult stem cell properties and for regenerative therapy approaches.

    Learn how to culture human intestinal organoids in our On-Demand Intestinal Course or browse our Frequently Asked Questions (FAQs) about the organoid workflows using IntestiCult™. Additionally, download our detailed e-book Proven Protocols for Intestinal Organoid Culture: Getting Started with IntestiCult™ for a curated collection of intestinal organoid protocols.

    Should you intend to use this product for commercial purposes, please contact HUB Organoids B.V. at www.huborganoids.nl for a commercial use license or for clarifications in relation to HUB Organoids B.V. licensing.
    Subtype
    Specialized Media
    Cell Type
    Intestinal Cells
    Species
    Human
    Application
    Cell Culture, Differentiation, Expansion, Maintenance, Organoid Culture
    Brand
    IntestiCult
    Area of Interest
    Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Stem Cell Biology

    Data Figures

    Figure 1. Primary Organoids Grown in IntestiCult™ Organoid Growth Medium (Human) are Fully Mature After 10-14 Days in Culture

    Primary organoids were cultured from human colonic biopsy samples and grown in IntestiCult Organoid Growth Medium (Human). Organoids were imaged after (A) two days, (B) six days, (C) eight days and (D) ten days growth.

    Figure 2. Organoids Grown in IntestiCult™ Organoid Growth Medium (Human) Display Markers of Human Intestinal Epithelial Cells

    Immunofluorescence of organoids grown in IntestiCult™ Organoid Growth Medium (Human) showing colocalization of (A) DAPI, (B) EPCAM and (C) Ki67. (D) A merged image shows the position of actively proliferating (Ki67+) intestinal stem cells within the epithelial layer (EPCAM+).

    Figure 3. Forskolin-Induced Swelling of Organoids Grown in IntestiCult™ Organoid Growth Medium (Human)

    Organoids were treated with (A) 5 μM Forskolin or (B) DMSO and organoid area was measured at 0 minutes and 120 minutes. (C)Forskolin-treated organoids increased in size 33.5 ± 3.8% compared to 7.5 ± 0.8% for DMSO-treated organoids.

    Figure 4. IntestiCult™ Organoid Growth Medium (Human) Supports the Growth of Organoids in Multiple Extracellular Matrices

    Intestinal organoid cultures were prepared in IntestiCult™ Organoid Growth Medium (Human) and plated in (A) Matrigel® Growth Factor Reduced Basement Membrane Matrix (Corning® catalog # 356231), (B) Geltrex® LDEV-Free Reduced Growth Factor Basement Membrane Matrix (Gibco™ catalog # A1413202), (C) Cultrex® Reduced Growth Factor Basement Membrane Extract, Type 1 (R&D Systems™ catalog # 3433-005-R1), and (D) Cultrex® Reduced Growth Factor Basement Membrane Extract, Type 2 (R&D Systems™ catalog # 3533-005-02). Organoid cultures are imaged at the end of passage 4. All four extracellular matrices supported robust growth of human intestinal organoids. Scale bars = 250 μm.

    Protocols and Documentation

    Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

    Document Type
    Product Name
    Catalog #
    Lot #
    Language
    Document Type
    Product Information Sheet
    Product Name
    IntestiCult™ Organoid Growth Medium (Human)
    Catalog #
    06010
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 1
    Product Name
    IntestiCult™ Organoid Growth Medium (Human)
    Catalog #
    06010
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 2
    Product Name
    IntestiCult™ Organoid Growth Medium (Human)
    Catalog #
    06010
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 3
    Product Name
    IntestiCult™ Organoid Growth Medium (Human)
    Catalog #
    06010
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 4
    Product Name
    IntestiCult™ Organoid Growth Medium (Human)
    Catalog #
    06010
    Lot #
    All
    Language
    English

    Applications

    This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

    Research Area
    Workflow Stages
    Workflow Stages for Organoids
    View All
    Workflow Stages for Genome Editing
    View All

    Resources and Publications

    Educational Materials (61)

    Intestinal Organoids
    Brochure
    Intestinal Organoids
    Organoids
    Brochure
    Organoids
    Culturing Cancer-Derived Organoids Using IntestiCult™ Organoid Growth Medium (Human)
    Technical Bulletin
    Culturing Cancer-Derived Organoids Using IntestiCult™ Organoid Growth Medium (Human)
    Co-Culturing Colorectal Organoids and T Cells using IntestiCult™ and ImmunoCult™
    Technical Bulletin
    Co-Culturing Colorectal Organoids and T Cells using IntestiCult™ and ImmunoCult™
    Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
    Technical Bulletin
    Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
    CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
    Protocol
    CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
    Culturing Intestinal Cancer-Derived Organoids
    Protocol
    Culturing Intestinal Cancer-Derived Organoids
    How to Isolate Antigen-Specific T Cells
    Protocol
    How to Isolate Antigen-Specific T Cells
    How to Perform Immunocytochemistry (ICC) Staining of Epithelial Cells Cultured as Monolayers or at the Air-Liquid Interface
    Protocol
    How to Perform Immunocytochemistry (ICC) Staining of Epithelial Cells Cultured as Monolayers or at t...
    How to Isolate Human Colonic Crypts from Intestinal Biopsies
    Protocol
    How to Isolate Human Colonic Crypts from Intestinal Biopsies
    How to Cryopreserve Intestinal Organoids
    Protocol
    How to Cryopreserve Intestinal Organoids
    Passaging Intestinal Organoids into Suspension Culture Using IntestiCult™
    Protocol
    Passaging Intestinal Organoids into Suspension Culture Using IntestiCult™
    Barrier Permeability Assay Using Human Intestinal Organoid-Derived Monolayers
    Protocol
    Barrier Permeability Assay Using Human Intestinal Organoid-Derived Monolayers
    Performing Immunocytochemical Staining of Epithelial Organoids
    Protocol
    Performing Immunocytochemical Staining of Epithelial Organoids
    How to Thaw Intestinal Organoids
    Protocol
    How to Thaw Intestinal Organoids
    How to Process Epithelial Organoids and Organoid-Derived Epithelial Monolayers for RNA Isolation
    Protocol
    How to Process Epithelial Organoids and Organoid-Derived Epithelial Monolayers for RNA Isolation
    SnapShot: GI Tract Development
    Wallchart
    SnapShot: GI Tract Development
    SnapShot: The Intestinal Crypt
    Wallchart
    SnapShot: The Intestinal Crypt
    SnapShot: Growing Organoids from Stem Cells
    Wallchart
    SnapShot: Growing Organoids from Stem Cells
    What Are the Impacts of New Legislative Actions on Drug Development?
    3:07
    Video
    What Are the Impacts of New Legislative Actions on Drug Development?
    Collaborating to Accelerate COVID-19 Research
    3:36
    Video
    Collaborating to Accelerate COVID-19 Research
    Growth of Mouse Pancreatic Organoids in PancreaCult™ Culture Medium: Under a Microscope
    1:07
    Video
    Growth of Mouse Pancreatic Organoids in PancreaCult™ Culture Medium: Under a Microscope
    How to Count Intestinal Crypts for Mouse Organoid Cultures
    1:09
    Video
    How to Count Intestinal Crypts for Mouse Organoid Cultures
    Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
    3:19
    Video
    Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
    Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
    56:31
    Webinar
    Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
    Challenges in Translating iPSC Technology
    41:47
    Webinar
    Challenges in Translating iPSC Technology
    Optimizing Organoid Models to Better Predict Clinical Outcomes
    55:09
    Webinar
    Optimizing Organoid Models to Better Predict Clinical Outcomes
    Modeling Host-Microbe Interactions Using Human Intestinal Organoids
    16:31
    Webinar
    Modeling Host-Microbe Interactions Using Human Intestinal Organoids
    Bioelectronic Tools to Study the Gut-Brain Axis
    40:52
    Webinar
    Bioelectronic Tools to Study the Gut-Brain Axis
    Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
    17:20
    Webinar
    Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
    Organoid Expert Panel
    52:35
    Webinar
    Organoid Expert Panel
    Organoid-Based Assays for Disease Modeling
    24:55
    Webinar
    Organoid-Based Assays for Disease Modeling
    Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
    4:47
    Webinar
    Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
    Advances in GI Organoid Applications: Drug Discovery and Immunotherapy
    47:49
    Webinar
    Advances in GI Organoid Applications: Drug Discovery and Immunotherapy
    Nature Research Round Table: Organoids as Models for Human Disease
    15:10
    Webinar
    Nature Research Round Table: Organoids as Models for Human Disease
    Patient-Derived Organoids for Drug Screening and Development
    50:25
    Webinar
    Patient-Derived Organoids for Drug Screening and Development
    GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
    24:56
    Webinar
    GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
    Elucidating Parechovirus A Pathogenesis Using Organoid Models
    30:09
    Webinar
    Elucidating Parechovirus A Pathogenesis Using Organoid Models
    Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
    15:15
    Webinar
    Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
    Dissecting the Innate Immune Response in Human Intestinal Organoid Models
    55:56
    Webinar
    Dissecting the Innate Immune Response in Human Intestinal Organoid Models
    Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
    10:17
    Webinar
    Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
    Applications of Human Intestinal Organoid Cultures in Drug Discovery
    56:20
    Webinar
    Applications of Human Intestinal Organoid Cultures in Drug Discovery
    HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
    1:10:09
    Webinar
    HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
    Using Stem Cell-Based Models in Drug Discovery
    46:49
    Webinar
    Using Stem Cell-Based Models in Drug Discovery
    ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases, Nephrotoxicity, and Highly Relevant Cell-Based Assay
    59:52
    Webinar
    ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases,...
    Molecular Tricks of Neurotropic Intestinal Viruses
    25:54
    Webinar
    Molecular Tricks of Neurotropic Intestinal Viruses
    Spotlight on Organoids: Expert Panel
    Webinar
    Spotlight on Organoids: Expert Panel
    Nature Research Round Table: The Promise of Organoid Medicine
    17:17
    Webinar
    Nature Research Round Table: The Promise of Organoid Medicine
    Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Development
    11:52
    Webinar
    Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Developm...
    Thinking Outside the Dome: Versatile Organotypic Cell Culture Methods
    39:57
    Webinar
    Thinking Outside the Dome: Versatile Organotypic Cell Culture Methods
    Adding Hepatic Organoids to Your Research
    50:59
    Webinar
    Adding Hepatic Organoids to Your Research
    Applications and Analysis of Organoids
    47:16
    Webinar
    Applications and Analysis of Organoids
    Intestinal Organoid Culture
    Mini Review
    Intestinal Organoid Culture
    Human Intestinal Organoid Culture System for Drug-Induced Gastrointestinal Toxicity Screening
    Scientific Poster
    Human Intestinal Organoid Culture System for Drug-Induced Gastrointestinal Toxicity Screening
    Highly Efficient Differentiation of Human Pluripotent Stem Cells into Long-term Expandable “Mini-gut” Organoids
    Scientific Poster
    Highly Efficient Differentiation of Human Pluripotent Stem Cells into Long-term Expandable “Mini-g...
    Tamara Zietek, PhD
    Interview
    Tamara Zietek, PhD
    Caroline Lindemans, MD, PhD
    Interview
    Caroline Lindemans, MD, PhD
    Dr. Bas Trietsch
    Interview
    Dr. Bas Trietsch
    Heather McCauley, PhD
    Interview
    Heather McCauley, PhD
    Jason Spence
    Interview
    Jason Spence
    Proven Protocols for Intestinal Organoid Culture: Getting Started with IntestiCult™
    E-Book
    Proven Protocols for Intestinal Organoid Culture: Getting Started with IntestiCult™

    Publications (26)

    In vivo development of immune tissue in human intestinal organoids transplanted into humanized mice. C. Bouffi et al. Nature biotechnology 2023 6

    Abstract

    Human intestinal organoids (HIOs) derived from pluripotent stem cells provide a valuable model for investigating human intestinal organogenesis and physiology, but they lack the immune components required to fully recapitulate the complexity of human intestinal biology and diseases. To address this issue and to begin to decipher human intestinal-immune crosstalk during development, we generated HIOs containing immune cells by transplanting HIOs under the kidney capsule of mice with a humanized immune system. We found that human immune cells temporally migrate to the mucosa and form cellular aggregates that resemble human intestinal lymphoid follicles. Moreover, after microbial exposure, epithelial microfold cells are increased in number, leading to immune cell activation determined by the secretion of IgA antibodies in the HIO lumen. This in vivo HIO system with human immune cells provides a framework for future studies on infection- or allergen-driven intestinal diseases.
    View publication
    Catechol-O-Methyltransferase Loss Drives Cell-Specific Nociceptive Signaling via the Enteric Catechol-O-Methyltransferase/microRNA-155/Tumor Necrosis Factor ? Axis Q. Zhou et al. Gastroenterology 2023 4

    Abstract

    BACKGROUND & AIMS The etiology of abdominal pain in postinfectious, diarrhea-predominant irritable bowel syndrome (PI-IBS-D) is unknown, and few treatment options exist. Catechol-O-methyltransferase (COMT), an enzyme that inactivates and degrades biologically active catecholamines, plays an important role in numerous physiologic processes, including modulation of pain perception. Our objective was to determine the mechanism(s) of how decreased colonic COMT in PI-IBS-D patients contributes to the chronic abdominal pain phenotype after enteric infections. METHODS Colon neurons, epithelial cells, and macrophages were procured with laser capture microdissection from PI-IBS-D patients to evaluate cell-specific colonic COMT, microRNA-155 (miR-155), and tumor necrosis factor (TNF) ? expression levels compared to recovered patients (infection cleared: did not develop PI-IBS-D) and control individuals. COMT-/-, colon-specific COMT-/-, and miR-155-/- mice and human colonoids were used to model phenotypic expression of COMT in PI-IBS-D patients and to investigate signaling pathways linking abdominal pain. Citrobacter rodentium and trinitrobenzene sulfonic acid animal models were used to model postinflammatory changes seen in PI-IBS-D patients. RESULTS Colonic COMT levels were significantly decreased and correlated with increased visual analog scale abdominal pain ratings in PI-IBS-D patients compared to recovered patients and control individuals. Colonic miR-155 and TNF-? were increased in PI-IBS-D patients with diminished colonic COMT. COMT-/- mice had significantly increased expression of miR-155 and TNF-? in both colon tissues and dorsal root ganglia. Introduction of cV1q antibody (anti-TNF-?) into mice reversed visceral hypersensitivity after C rodentium and trinitrobenzene sulfonic acid. CONCLUSIONS Decreased colonic COMT in PI-IBS-D patients drives abdominal pain phenotypes via the COMT/miR-155/TNF-? axis. These important findings will allow new treatment paradigms and more targeted and personalized medicine approaches for gastrointestinal disorders after enteric infections.
    View publication
    3D bioprinted colorectal cancer models based on hyaluronic acid and signalling glycans. F. Cadamuro et al. Carbohydrate polymers 2023 2

    Abstract

    In cancer microenvironment, aberrant glycosylation events of ECM proteins and cell surface receptors occur. We developed a protocol to generate 3D bioprinted models of colorectal cancer (CRC) crosslinking hyaluronic acid and gelatin functionalized with three signalling glycans characterized in CRC, 3'-Sialylgalactose, 6'-Sialylgalactose and 2'-Fucosylgalactose. The crosslinking, performed exploiting azide functionalized gelatin and hyaluronic acid and 4arm-PEG-dibenzocyclooctyne, resulted in biocompatible hydrogels that were 3D bioprinted with commercial CRC cells HT-29 and patient derived CRC tumoroids. The glycosylated hydrogels showed good 3D printability, biocompatibility and stability over the time. SEM and synchrotron radiation SAXS/WAXS analysis revealed the influence of glycosylation in the construct morphology, whereas MALDI-MS imaging showed that protein profiles of tumoroid cells vary with glycosylation, indicating that sialylation and fucosylation of ECM proteins induce diverse alterations to the proteome of the tumoroid and surrounding cells.
    View publication
    View All Publications

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    Legal Statement:

    This product was developed under a license to intellectual property owned by Hubrecht Organoid Technology (HUB). This product is sold for research use only. Purchase of this product does not include the right to use it for drug screening aiming for commercial gain, equipment validation, biobanking, or for other commercial purposes. Purchasers wishing to use the product for purposes other than basic research use should contact HUB at www.huborganoids.nl to obtain a further license. Purchasers may apply for a License from HUB, which will not be unreasonably withheld by HUB.

    Quality Statement:

    PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.

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