IntestiCult™ Organoid Growth Medium (Human)
Cell culture medium for establishment and maintenance of human intestinal organoids
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Required Products
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Gentle Cell Dissociation ReagentcGMP, enzyme-free cell dissociation reagent
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DMEM/F-12 with 15 mM HEPESDulbecco's Modified Eagle's Medium/Nutrient Ham's Mixture F-12 (DMEM/F-12) with 15 mM HEPES buffer
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Y-27632RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
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D-PBS (Without Ca++ and Mg++)Dulbecco’s phosphate-buffered saline without calcium and magnesium
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.
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.
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
• Complete medium formulation that delivers consistent results
• Enables generation of intestinal organoids in one week
• Easy-to-use format and optimized protocol
Components
- IntestiCult™ OGM Human Basal Medium, 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
Related Products
Related Products
Scientific Resources
Product Documentation
| Document Type | Product Name | Catalog # | Lot # | Language |
|---|---|---|---|---|
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Document Type
Product Information Sheet
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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 |
Educational Materials (44)
Brochure
Intestinal Organoids
Brochure
Organoids
Brochure
2019-2020 Cell Culture Training Catalog
Technical Bulletin
Culturing Cancer-Derived Organoids Using IntestiCult™ Organoid Growth Medium (Human)
Technical Bulletin
Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
Protocol
CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
Protocol
How to Generate Mouse Intestinal Organoid-Derived Monolayers Using IntestiCult™
Protocol
Performing Immunocytochemical Staining of Epithelial Organoids
Wallchart
SnapShot: GI Tract Development
Wallchart
SnapShot: Growing Organoids from Stem Cells
Wallchart
SnapShot: The Intestinal Crypt
Video
1:09
How to Count Intestinal Crypts for Mouse Organoid Cultures
Video
3:19
Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
Video
3:36
Collaborating to Accelerate COVID-19 Research
Video
1:07
Growth of Mouse Pancreatic Organoids in PancreaCult™ Culture Medium: Under a Microscope
Video
1:18
Growth of Liver Organoids in HepatiCult™ Culture Medium: Under a Microscope
Webinar
41:47
Challenges in Translating iPSC Technology
Webinar
52:35
Organoid Expert Panel
Webinar
17:20
Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
Webinar
56:31
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
Webinar
4:47
Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
Webinar
9:36
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
Webinar
24:56
GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
Webinar
Spotlight on Organoids: Expert Panel
Webinar
1:10:09
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
Webinar
50:25
Patient-Derived Organoids for Drug Screening and Development
Webinar
15:15
Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
Webinar
11:52
Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Development
Webinar
15:10
Nature Research Round Table: Organoids as Models for Human Disease
Webinar
16:31
Modeling Host-Microbe Interactions Using Human Intestinal Organoids
Webinar
59:52
ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases, Nephrotoxicity, and Highly Relevant Cell-Based Assay
Webinar
50:59
Adding Hepatic Organoids to Your Research
Webinar
17:17
Nature Research Round Table: The Promise of Organoid Medicine
Webinar
12:57
Nature Research Round Table: Liver Organoids for the Study of Liver Regeneration and Disease
Webinar
10:17
Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
Mini Review
Intestinal Organoid Culture
Scientific Poster
Modelling Pancreatic Cancer Through Pancreatic Exocrine Organoids Using PancreaCult™ Serum-free Medium
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)
Scientific Poster
Highly Efficient Differentiation of Human Pluripotent Stem Cells into Long-term Expandable “Mini-gut” Organoids
Interview
Caroline Lindemans, MD, PhD
Interview
Luigi Aloia, PhD
Interview
Jason Spence
Interview
Heather McCauley, PhD
Interview
Tamara Zietek, PhD
Load More Educational Materials
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
Workflow Stages for
Intestinal Epithelial Cell Research
Workflow Stages for
Genome Editing
Data and Publications
Data
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.
Publications (18)
ACS infectious diseases 2020 may
Fucose-Galactose Polymers Inhibit Cholera Toxin Binding to Fucosylated Structures and Galactose-Dependent Intoxication of Human Enteroids.
Abstract
Abstract
A promising strategy to limit cholera severity involves blockers mimicking the canonical cholera toxin ligand (CT) ganglioside GM1. However, to date the efficacies of most of these blockers have been evaluated in noncellular systems that lack ligands other than GM1. Importantly, the CT B subunit (CTB) has a noncanonical site that binds fucosylated structures, which in contrast to GM1 are highly expressed in the human intestine. Here we evaluate the capacity of norbornene polymers displaying galactose and/or fucose to block CTB binding to immobilized protein-linked glycan structures and also to primary human and murine small intestine epithelial cells (SI ECs). We show that the binding of CTB to human SI ECs is largely dependent on the noncanonical binding site, and interference with the canonical site has a limited effect while the opposite is observed with murine SI ECs. The galactose-fucose polymer blocks binding to fucosylated glycans but not to GM1. However, the preincubation of CT with the galactose-fucose polymer only partially blocks toxic effects on cultured human enteroid cells, while preincubation with GM1 completely blocks CT-mediated secretion. Our results support a model whereby the binding of fucose to the noncanonical site places CT in close proximity to scarcely expressed galactose receptors such as GM1 to enable binding via the canonical site leading to CT internalization and intoxication. Our finding also highlights the importance of complementing CTB binding studies with functional intoxication studies when assessing the efficacy inhibitors of CT.
Nature communications 2020 may
TNFAIP8 controls murine intestinal stem cell homeostasis and regeneration by regulating microbiome-induced Akt signaling.
Abstract
Abstract
The intestine is a highly dynamic environment that requires tight control of the various inputs to maintain homeostasis and allow for proper responses to injury. It was recently found that the stem cell niche and epithelium is regenerated after injury by de-differentiated adult cells, through a process that gives rise to Sca1+ fetal-like cells and is driven by a transient population of Clu+ revival stem cells (revSCs). However, the molecular mechanisms that regulate this dynamic process have not been fully defined. Here we show that TNFAIP8 (also known as TIPE0) is a regulator of intestinal homeostasis that is vital for proper regeneration. TIPE0 functions through inhibiting basal Akt activation by the commensal microbiota via modulating membrane phospholipid abundance. Loss of TIPE0 in mice results in injury-resistant enterocytes, that are hyperproliferative, yet have regenerative deficits and are shifted towards a de-differentiated state. Tipe0-/- enterocytes show basal induction of the Clu+ regenerative program and a fetal gene expression signature marked by Sca1, but upon injury are unable to generate Sca-1+/Clu+ revSCs and could not regenerate the epithelium. This work demonstrates the role of TIPE0 in regulating the dynamic signaling that determines the injury response and enables intestinal epithelial cell regenerative plasticity.
Toxicology in vitro : an international journal published in association with BIBRA 2020 jul
Human ileal organoid model recapitulates clinical incidence of diarrhea associated with small molecule drugs.
Abstract
Abstract
Drug-induced gastrointestinal toxicity (GIT) is a common treatment-emergent adverse event that can negatively impact dosing, thereby limiting efficacy and treatment options for patients. An in vitro assay of GIT is needed to address patient variability, mimic the microphysiology of the gut, and accurately predict drug-induced GIT. Primary human ileal organoids (termed 'enteroids') have proven useful for stimulating intestinal stem cell proliferation and differentiation to multiple cell types present in the gut epithelium. Enteroids have enabled characterization of gut biology and the signaling involved in the pathogenesis of disease. Here, enteroids were differentiated from four healthy human donors and assessed for culture duration-dependent differentiation status by immunostaining for gut epithelial markers lysozyme, chromogranin A, mucin, and sucrase isomaltase. Differentiated enteroids were evaluated with a reference set of 31 drugs exhibiting varying degrees of clinical incidence of diarrhea, a common manifestation of GIT that can be caused by drug-induced thinning of the gut epithelium. An assay examining enteroid viability in response to drug treatment demonstrated 90{\%} accuracy for recapitulating the incidence of drug-induced diarrhea. The human enteroid viability assay developed here presents a promising in vitro model for evaluating drug-induced diarrhea.
The Journal of clinical investigation 2020 jan
Apelin directs endothelial cell differentiation and vascular repair following immune-mediated injury.
Abstract
Abstract
Sustained, indolent immune injury of the vasculature of a heart transplant limits long-term graft and recipient survival. This injury is mitigated by a poorly characterized, maladaptive repair response. Vascular endothelial cells respond to proangiogenic cues in the embryo by differentiation to specialized phenotypes, associated with expression of apelin. In the adult, the role of developmental proangiogenic cues in repair of the established vasculature is largely unknown. We found that human and minor histocompatibility-mismatched donor mouse heart allografts with alloimmune-mediated vasculopathy upregulated expression of apelin in arteries and myocardial microvessels. In vivo, loss of donor heart expression of apelin facilitated graft immune cell infiltration, blunted vascular repair, and worsened occlusive vasculopathy in mice. In vitro, an apelin receptor agonist analog elicited endothelial nitric oxide synthase activation to promote endothelial monolayer wound repair and reduce immune cell adhesion. Thus, apelin acted as an autocrine growth cue to sustain vascular repair and mitigate the effects of immune injury. Treatment with an apelin receptor agonist after vasculopathy was established markedly reduced progression of arterial occlusion in mice. Together, these initial data identify proangiogenic apelin as a key mediator of coronary vascular repair and a pharmacotherapeutic target for immune-mediated injury of the coronary vasculature.
Journal of animal science 2020 feb
Evaluation of swine enteroids as in vitro models for Lawsonia intracellularis infection1,2.
Abstract
Abstract
The enteric pathogen Lawsonia intracellularis is one of the main causes of diarrhea and compromised weight gain in pigs worldwide. Traditional cell-line cultures have been used to study L. intracellularis pathogenesis. However, these systems fail to reproduce the epithelial changes observed in the intestines of L. intracellularis-infected pigs, specifically, the changes in intestinal cell constitution and gene expression. A more physiologically accurate and state-of-the-art model is provided by swine enteroids derived from stem cell-containing crypts from healthy pigs. The objective of this study was to verify the feasibility of two-dimensional swine enteroids as in vitro models for L. intracellularis infection. We established both three- and two-dimensional swine enteroid cultures derived from intestinal crypts. The two-dimensional swine enteroids were infected by L. intracellularis in four independent experiments. Enteroid-infected samples were collected 3 and 7 d postinfection for analysis using real-time quantitative PCR and L. intracellularis immunohistochemistry. In this study, we show that L. intracellularis is capable of infecting and replicating intracellularly in two-dimensional swine enteroids derived from ileum.
mBio 2020
Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection.
Abstract
Abstract
Human noroviruses (HuNoVs) are the leading cause of nonbacterial gastroenteritis worldwide. Histo-blood group antigen (HBGA) expression is an important susceptibility factor for HuNoV infection based on controlled human infection models and epidemiologic studies that show an association of secretor status with infection caused by several genotypes. The fucosyltransferase 2 gene (FUT2) affects HBGA expression in intestinal epithelial cells; secretors express a functional FUT2 enzyme, while nonsecretors lack this enzyme and are highly resistant to infection and gastroenteritis caused by many HuNoV strains. These epidemiologic associations are confirmed by infections in stem cell-derived human intestinal enteroid (HIE) cultures. GII.4 HuNoV does not replicate in HIE cultures derived from nonsecretor individuals, while HIEs from secretors are permissive to infection. However, whether FUT2 expression alone is critical for infection remains unproven, since routinely used secretor-positive transformed cell lines are resistant to HuNoV replication. To evaluate the role of FUT2 in HuNoV replication, we used CRISPR or overexpression to genetically manipulate FUT2 gene function to produce isogenic HIE lines with or without FUT2 expression. We show that FUT2 expression alone affects both HuNoV binding to the HIE cell surface and susceptibility to HuNoV infection. These findings indicate that initial binding to a molecule(s) glycosylated by FUT2 is critical for HuNoV infection and that the HuNoV receptor is present in nonsecretor HIEs. In addition to HuNoV studies, these isogenic HIE lines will be useful tools to study other enteric microbes where infection and/or disease outcome is associated with secretor status.IMPORTANCE Several studies have demonstrated that secretor status is associated with susceptibility to human norovirus (HuNoV) infection; however, previous reports found that FUT2 expression is not sufficient to allow infection with HuNoV in a variety of continuous laboratory cell lines. Which cellular factor(s) regulates susceptibility to HuNoV infection remains unknown. We used genetic manipulation of HIE cultures to show that secretor status determined by FUT2 gene expression is necessary and sufficient to support HuNoV replication based on analyses of isogenic lines that lack or express FUT2. Fucosylation of HBGAs is critical for initial binding and for modification of another putative receptor(s) in HIEs needed for virus uptake or uncoating and necessary for successful infection by GI.1 and several GII HuNoV strains.
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 this product for growing organoids for drug screening aiming for commercial gain or for other commercial purposes. Purchasers wishing to use the product for purposes other than research use should contact HUB to obtain a further license (an “Organoid-Growth License”). Purchasers may apply for an Organoid-Growth License from HUB and such license 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.
