IntestiCult™ Organoid Growth Medium (Human)

Cell culture medium for establishment and maintenance of human intestinal organoids
IntestiCult™ Organoid Growth Medium (Human)

Cell culture medium for establishment and maintenance of human intestinal organoids

1 Kit
Catalog # 06010
Required Products
  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

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

Overview

IntestiCult™ Organoid Growth Medium (Human) is a complete cell culture medium for efficient establishment and long-term maintenance of intestinal organoids derived from human intestinal crypts. Intestinal organoids provide a convenient in vitro organotypic culture system for studying the intestinal epithelium. The organoids incorporate a functional lumen enclosed by a polarized intestinal epithelial cell layer. Isolated intestinal crypts rapidly form organoids when cultured in IntestiCult™ Organoid Growth Medium (Human). 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
Components
  • IntestiCult™ OGM Human Basal Medium
  • Organoid Supplement
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

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet 1
Product Name
IntestiCult™ Organoid Growth Medium (Human)
Catalog #
06010
Lot #
Purchase date 2020-08-05 or later.
Language
English
Document Type
Product Information Sheet 2
Product Name
IntestiCult™ Organoid Growth Medium (Human)
Catalog #
06010
Lot #
Component #06011: Lot 17E80031
Language
English
Document Type
Product Information Sheet 3
Product Name
IntestiCult™ Organoid Growth Medium (Human)
Catalog #
06010
Lot #
Component #06011: Lot 18B88629 or higher
Language
English
Document Type
Special Protocol 1
Product Name
IntestiCult™ Organoid Growth Medium (Human)
Catalog #
06010
Lot #
All
Language
English
Document Type
Special Protocol 2
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

Educational Materials(41)

Brochure
Organoids
Brochure
2019-2020 Cell Culture Training Catalog
Brochure
Intestinal Organoids
Technical Bulletin
Culturing Cancer-Derived Organoids Using IntestiCult™ Organoid Growth Medium (Human)
Technical Bulletin
Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
Wallchart
SnapShot: Growing Organoids from Stem Cells
Wallchart
SnapShot: The Intestinal Crypt
Wallchart
SnapShot: GI Tract Development
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
Video
1:09
How to Count Intestinal Crypts for Mouse Organoid Cultures
Video
3:19
Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
Webinar
41:47
Challenges in Translating iPSC Technology
Webinar
52:35
Organoid Expert Panel
Webinar
56:31
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
Webinar
17:20
Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
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
50:25
Patient-Derived Organoids for Drug Screening and Development
Webinar
1:10:09
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
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
50:59
Adding Hepatic Organoids to Your Research
Webinar
59:52
ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases, Nephrotoxicity, and Highly Relevant Cell-Based Assay
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
Heather McCauley, PhD
Interview
Tamara Zietek, PhD
Interview
Caroline Lindemans, MD, PhD
Interview
Luigi Aloia, PhD
Interview
Jason Spence
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

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)

Nature communications 2020 may TNFAIP8 controls murine intestinal stem cell homeostasis and regeneration by regulating microbiome-induced Akt signaling. J. R. Goldsmith et al.

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.
ACS infectious diseases 2020 may Fucose-Galactose Polymers Inhibit Cholera Toxin Binding to Fucosylated Structures and Galactose-Dependent Intoxication of Human Enteroids. J. Cervin et al.

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.
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. D. G. Belair et al.

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. A. G. Masoud et al.

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. T. P. Resende et al.

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.
Frontiers in cellular and infection microbiology 2020 A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine. T. Roodsant et al.

Abstract

Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and Listeria monocytogenes were evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research.
View All Publications

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