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IntestiCult™ Organoid Differentiation Medium (Human)

Cell culture medium for further differentiation of human intestinal organoids in 3D, or as monolayers/air-liquid interface cultures

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IntestiCult™ Organoid Differentiation Medium (Human)

Cell culture medium for further differentiation of human intestinal organoids in 3D, or as monolayers/air-liquid interface cultures

From: 317 USD
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Cell culture medium for further differentiation of human intestinal organoids in 3D, or as monolayers/air-liquid interface cultures
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Product Advantages


  • Enables generation of intestinal organoid cultures with physiological proportions of differentiated and stem cell populations

  • Allows easy access to the apical surface by transitioning intestinal organoids into monolayer and ALI cultures

  • Generates consistent results across passages and replicates

What's Included

  • IntestiCult™ ODM 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. IntestiCult™ Organoid Growth Medium (Human)
    IntestiCult™ Organoid Growth Medium (Human)

    Cell culture medium for establishment and maintenance of human intestinal organoids

  2. Gentle Cell Dissociation Reagent
    Gentle Cell Dissociation Reagent

    cGMP, enzyme-free cell dissociation reagent

  3. DAPT
    DAPT

    Notch pathway inhibitor; Inhibits γ-secretase

  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

IntestiCult™ Organoid Differentiation Medium (Human) is a complete culture medium that supports the further differentiation of intestinal organoids in three dimensions (3D), or in 2D as monolayers or air-liquid interface (ALI) cultures. The starting cultures can be either intestinal organoids derived from human intestinal crypts, or passaged organoids that have been cultured with IntestiCult™ Organoid Growth Medium (Human; Catalog #06010).

Intestinal cultures generated using IntestiCult™ Organoid Differentiation Medium (Human) contain physiologically relevant proportions of differentiated and stem cell populations, recapitulating the diversity of the crypt-villus axis. When compared to conventional cell lines, intestinal monolayers exhibit greater barrier integrity, express higher levels of key differentiation markers, and have a morphology that is more representative of the in vivo intestine.

Applications of intestinal organoid cultures include studying the development and function of the intestinal epithelium, modeling intestinal diseases, compound screening, and regenerative therapy approaches. Intestinal monolayer and ALI cultures are particularly amenable for permeability assays and studies of infectious diseases due to easy access to the apical surface. This kit requires IntestiCult™ Organoid Growth Medium (Human; Catalog #06010) for the initiation and expansion of intestinal organoids prior to differentiation.

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

Differentiated Human Intestinal Organoids Display a Budded Morphology

Figure 1. Differentiated Human Intestinal Organoids Display a Budded Morphology

(A) Organoids grown in IntestiCult™ OGM are primarily cystic. (B) When switched to IntestiCult™ ODM, organoids develop a thickened epithelium with a pronounced, budded morphology indicative of a more differentiated state. Organoids were imaged on day 5 of expansion or differentiation respectively.

Intestinal Organoids Contain a Higher Proportion of Mature Cell Types Following Differentiation in IntestiCult™ ODM

Figure 2. Intestinal Organoids Contain a Higher Proportion of Mature Cell Types Following Differentiation in IntestiCult™ ODM

(A, B) Organoids grown in IntestiCult™ OGM are enriched for Ki-67+ proliferative cells (A), while containing few differentiated cell types such as goblet cells (MUC2, A), enterocytes (KRT20, B), and enteroendocrine cells (CHGA, B). (C, D) When switched to IntestiCult™ ODM, organoids contain a small number of Ki-67+ proliferative cells (C, arrows), with more physiological proportions of goblet cells (MUC2, C), enterocytes (KRT20, D), and chromogranin A- (CHGA-)positive enteroendocrine cells (D, arrow).

Differentiation of Intestinal Epithelium at the Air-Liquid Interface (ALI) Using IntestiCult™ ODM

Figure 3. Differentiation of Intestinal Epithelium at the Air-Liquid Interface (ALI) Using IntestiCult™ ODM

(A – E) Growing organoid-derived monolayers as ALI cultures drives further differentiation of intestinal epithelial cultures as seen by changes in gene expression measured by RT-qPCR. Relative quantification (RQ) for each marker is shown relative to actB and TBP housekeeping genes and normalized with respect to undifferentiated organoids grown in IntestiCult OGM (Human). Progenitor markers (A) Lgr5 and (B) Axin2 are significantly reduced in both submerged monolayers and ALI cultures, while markers of enterocytes (KRT20, C), goblet cells (MUC2, D), and enteroendocrine cells (CHGA, E) are significantly increased. Further reduction in Axin2 is seen in ALI monolayers with an increase in expression of KRT20, MUC2, and CHGA. (F, G) Comparing cross-sections of organoid monolayers grown in IntestiCult™ ODM as (F) submerged culture or (G) at the ALI shows further differentiation of the intestinal epithelium with an increased proportion of goblet cells and extracellular mucus (MUC2, green).

Differentiated Organoid-Derived Monolayers and ALI Cultures Display More Physiological Trans-Epithelial Electrical Resistance (TEER) than Caco-2 Cells

Figure 4. Differentiated Organoid-Derived Monolayers and ALI Cultures Display More Physiological Trans-Epithelial Electrical Resistance (TEER) than Caco-2 Cells

Differentiated organoid-derived monolayers grown as a submerged monolayer (IntestiCult™ ODM Monolayer), or at the ALI (IntestiCult™ ODM ALI), show higher TEER values as compared to Caco-2 cultures.Organoid-derived monolayers grown at the ALI show a loosening of tight junctions due to further differentiation of the brush border, and thus lower TEER values are observed. * p < 0.0001.

Differentiated Intestinal Organoids Provide a Suitable Model for Studying CFTR Response In Vitro

Figure 5. Differentiated Intestinal Organoids Provide a Suitable Model for Studying CFTR Response In Vitro

(A) Organoids differentiated further in IntestiCult™ ODM show a comparable degree of swelling when treated with forskolin as compared to organoids grown in IntestiCult™ OGM, demonstrating suitability for use in forskolin-induced swelling assays. (B – E) Ussing chamber analysis of submerged (B) organoid-derived monolayers and (C) Caco-2 cultures demonstrate increased sensitivity of organoid-derived monolayers to CFTR activation and inhibition by IBMX/Forskolin and CFTR Inhibitor-172 respectively. (D, E) Analysis of CFTR modulation by IBMX/Forskolin and CFTR Inhibitor-172 show significantly greater (D) activation and (E) inhibition of CFTR activity in organoid-derived monolayers as compared to Caco-2 cultures (p < 0.001 for both).

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 Differentiation Medium (Human)
Catalog #
100-0214
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
IntestiCult™ Organoid Differentiation Medium (Human)
Catalog #
100-0214
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
IntestiCult™ Organoid Differentiation Medium (Human)
Catalog #
100-0214
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

Resources and Publications

Educational Materials (51)

Organoids
Brochure
Organoids
Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
Technical Bulletin
Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
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
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...
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 Generate Monolayers from hPSC-Derived Organoids Using IntestiCult™
Protocol
How to Generate Monolayers from hPSC-Derived Organoids Using IntestiCult™
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
Culturing Intestinal Cancer-Derived Organoids
Protocol
Culturing Intestinal Cancer-Derived Organoids
CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
Protocol
CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
SnapShot: Growing Organoids from Stem Cells
Wallchart
SnapShot: Growing Organoids from Stem Cells
SnapShot: GI Tract Development
Wallchart
SnapShot: GI Tract Development
SnapShot: The Intestinal Crypt
Wallchart
SnapShot: The Intestinal Crypt
Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
3:19
Video
Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
How to Count Intestinal Crypts for Mouse Organoid Cultures
1:09
Video
How to Count Intestinal Crypts for Mouse Organoid Cultures
Collaborating to Accelerate COVID-19 Research
3:36
Video
Collaborating to Accelerate COVID-19 Research
How to Perform a TEER Measurement to Evaluate Epithelial Barrier Integrity in ALI Cultures
4:25
Video
How to Perform a TEER Measurement to Evaluate Epithelial Barrier Integrity in ALI Cultures
Correlating TEER Values with Air-Liquid Interface Culture Morphology
3:02
Video
Correlating TEER Values with Air-Liquid Interface Culture Morphology
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
Applications and Analysis of Organoids
47:16
Webinar
Applications and Analysis of Organoids
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
Applications of Human Intestinal Organoid Cultures in Drug Discovery
56:20
Webinar
Applications of Human Intestinal Organoid Cultures in Drug Discovery
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
Optimizing Organoid Models to Better Predict Clinical Outcomes
55:09
Webinar
Optimizing Organoid Models to Better Predict Clinical Outcomes
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
Organoid Expert Panel
52:35
Webinar
Organoid Expert Panel
Organoid Mini-Symposium: Human Intestinal Organoids to Study Rare Gut Endocrine Cells
Webinar
Organoid Mini-Symposium: Human Intestinal Organoids to Study Rare Gut Endocrine Cells
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: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
15:15
Webinar
Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
56:31
Webinar
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
9:36
Webinar
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
From Stem Cells to Organoids: Modeling the Gastrointestinal Tract
1:03:40
Webinar
From Stem Cells to Organoids: Modeling the Gastrointestinal Tract
Bioelectronic Tools to Study the Gut-Brain Axis
40:52
Webinar
Bioelectronic Tools to Study the Gut-Brain Axis
Challenges in Translating iPSC Technology
41:47
Webinar
Challenges in Translating iPSC Technology
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
1:10:09
Webinar
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
Organoid Mini-Symposium: Utilization of Next Generation CRISPR Tools to Model Health and Disease in Adult Stem Cell-Derived Organoids
Webinar
Organoid Mini-Symposium: Utilization of Next Generation CRISPR Tools to Model Health and Disease in ...
Patient-Derived Organoids for Drug Screening and Development
50:25
Webinar
Patient-Derived Organoids for Drug Screening and Development
Nature Research Round Table: The Promise of Organoid Medicine
17:17
Webinar
Nature Research Round Table: The Promise of Organoid Medicine
Intestinal Organoids: Improved Tools and In Vitro Models for Drug Development
17:32
Webinar
Intestinal Organoids: Improved Tools and In Vitro Models for Drug Development
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...
Nature Research Round Table: Organoids as Models for Human Disease
15:10
Webinar
Nature Research Round Table: Organoids as Models for Human Disease
Molecular Tricks of Neurotropic Intestinal Viruses
25:54
Webinar
Molecular Tricks of Neurotropic Intestinal Viruses
Organoid-Based Assays for Disease Modeling
24:55
Webinar
Organoid-Based Assays for Disease Modeling
Spotlight on Organoids: Expert Panel
Webinar
Spotlight on Organoids: Expert Panel
Organoid Mini-Symposium: Advanced Organoid Culture Systems and Their Applications in Infectious Diseases
32:21
Webinar
Organoid Mini-Symposium: Advanced Organoid Culture Systems and Their Applications in Infectious Dise...
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...
Efficient Establishment and Growth of Human Intestinal Organoid-Derived Monolayers
Scientific Poster
Efficient Establishment and Growth of Human Intestinal Organoid-Derived Monolayers
Dr. Bas Trietsch
Interview
Dr. Bas Trietsch
Proven Protocols for Intestinal Organoid Culture: Getting Started with IntestiCult™
E-Book
Proven Protocols for Intestinal Organoid Culture: Getting Started with IntestiCult™

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Interested in trying STEMCELL’s organoid products for your intestinal research? Fill out the form to request information about introductory offers.

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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