IntestiCult™ Organoid Differentiation Medium (Human)

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

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.

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)
1 Kit
288 USD
Catalog # 100-0214

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

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
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.
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

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™.
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
Catalog #
100-0214
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
100-0214
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
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.

Resources and Publications

Educational Materials (41)

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