IntestiCult™ Organoid Differentiation Medium (Human)

Cell culture medium for further differentiation of human intestinal organoids in 3D, or as monolayers/air-liquid interface cultures
IntestiCult™ Organoid Differentiation Medium (Human)

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

1 Kit
Catalog # 100-0214
264 USD
You may notice that your reagent packaging looks slightly different from images displayed here or from previous orders. Due to pandemic-related plasticware shortages, we are temporarily using alternative bottles for this product. Rest assured that the products themselves and how you should use them have not changed.
Required Products
  1. Gentle Cell Dissociation Reagent
    Gentle Cell Dissociation Reagent

    cGMP, enzyme-free cell dissociation reagent

  2. DAPT
    DAPT

    Notch pathway inhibitor; Inhibits γ-secretase

  3. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

  4. IntestiCult™ Organoid Growth Medium (Human)
    IntestiCult™ Organoid Growth Medium (Human)

    Cell culture medium for establishment and maintenance of human intestinal organoids

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.
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
Components
  • IntestiCult™ ODM Human Basal Medium, 50 mL
  • Organoid Supplement, 50 mL
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

Scientific Resources

Product Documentation

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

Educational Materials (39)

Brochure
Organoids
Technical Bulletin
Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
Protocol
How to Perform Immunocytochemistry (ICC) Staining of Epithelial Cells Cultured as Monolayers or at the Air-Liquid Interface
Protocol
How to Generate Mouse Intestinal Organoid-Derived Monolayers Using IntestiCult™
Protocol
Performing Immunocytochemical Staining of Epithelial Organoids
Protocol
CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
Wallchart
SnapShot: Growing Organoids from Stem Cells
Wallchart
SnapShot: The Intestinal Crypt
Wallchart
SnapShot: GI Tract Development
Video
How to Perform a TEER Measurement to Evaluate Epithelial Barrier Integrity in ALI Cultures
4:25
How to Perform a TEER Measurement to Evaluate Epithelial Barrier Integrity in ALI Cultures
Video
Correlating TEER Values with Air-Liquid Interface Culture Morphology
3:02
Correlating TEER Values with Air-Liquid Interface Culture Morphology
Video
How to Count Intestinal Crypts for Mouse Organoid Cultures
1:09
How to Count Intestinal Crypts for Mouse Organoid Cultures
Video
Collaborating to Accelerate COVID-19 Research
3:36
Collaborating to Accelerate COVID-19 Research
Webinar
Nature Research Round Table: The Promise of Organoid Medicine
17:17
Nature Research Round Table: The Promise of Organoid Medicine
Webinar
Nature Research Round Table: Liver Organoids for the Study of Liver Regeneration and Disease
12:57
Nature Research Round Table: Liver Organoids for the Study of Liver Regeneration and Disease
Webinar
Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
10:17
Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
Webinar
Challenges in Translating iPSC Technology
41:47
Challenges in Translating iPSC Technology
Webinar
Patient-Derived Organoids for Drug Screening and Development
50:25
Patient-Derived Organoids for Drug Screening and Development
Webinar
Organoid Mini-Symposium: Advanced Organoid Culture Systems and Their Applications in Infectious Diseases
32:21
Organoid Mini-Symposium: Advanced Organoid Culture Systems and Their Applications in Infectious Diseases
Webinar
Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
17:20
Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
Webinar
Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
4:47
Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
Webinar
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
9:36
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
Webinar
GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
24:56
GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
Webinar
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
1:10:09
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
Webinar
Organoid Expert Panel
52:35
Organoid Expert Panel
Webinar
From Stem Cells to Organoids: Modeling the Gastrointestinal Tract
1:03:40
From Stem Cells to Organoids: Modeling the Gastrointestinal Tract
Webinar
Organoid Mini-Symposium: Human Intestinal Organoids to Study Rare Gut Endocrine Cells
Webinar
Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
15:15
Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
Webinar
Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Development
11:52
Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Development
Webinar
Nature Research Round Table: Organoids as Models for Human Disease
15:10
Nature Research Round Table: Organoids as Models for Human Disease
Webinar
Intestinal Organoids: Improved Tools and In Vitro Models for Drug Development
17:32
Intestinal Organoids: Improved Tools and In Vitro Models for Drug Development
Webinar
Spotlight on Organoids: Expert Panel
Webinar
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
56:31
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
Webinar
Organoid Mini-Symposium: Utilization of Next Generation CRISPR Tools to Model Health and Disease in Adult Stem Cell-Derived Organoids
Scientific Poster
Highly Efficient Differentiation of Human Pluripotent Stem Cells into Long-term Expandable “Mini-gut” Organoids
Scientific Poster
Efficient Establishment and Long-Term Maintenance of 3-Dimensional Mouse Intestinal Organoids Using a Novel Defined and Serum-Free Medium
Scientific Poster
Efficient Establishment and Growth of Human Intestinal Organoid-Derived Monolayers
Scientific Poster
Development of a 96-well Assay for Assessing Cell Viability in Mouse Small Intestinal-Derived Organoids after Treatment with Cytotoxic Compounds
Scientific Poster
Efficient Establishment and Long-term Maintenance of 3-dimensional Human Small Intestinal and Colonic Organoids Using a Novel IntestiCult™ Organoid Growth Medium (Human)
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Product 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

Data and Publications

Data

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).

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