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
509 USD
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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. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

  4. D-PBS Without Ca++ and Mg++
    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.
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, 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 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
    Forskolin-Induced Swelling of Human Intestinal Organoids Grown in IntestiCult™
    Technical Bulletin
    Culturing Cancer-Derived Organoids Using IntestiCult™ Organoid Growth Medium (Human)
    Protocol
    Performing Immunocytochemical Staining of Epithelial Organoids
    Protocol
    CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
    Protocol
    How to Generate Mouse Intestinal Organoid-Derived Monolayers Using IntestiCult™
    Wallchart
    SnapShot: GI Tract Development
    Wallchart
    SnapShot: Growing Organoids from Stem Cells
    Wallchart
    SnapShot: The Intestinal Crypt
    Video
    How to Count Intestinal Crypts for Mouse Organoid Cultures
    1:09
    How to Count Intestinal Crypts for Mouse Organoid Cultures
    Video
    Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
    3:19
    Why Culture Intestinal Organoids with IntestiCult™ Organoid Growth Medium
    Video
    Collaborating to Accelerate COVID-19 Research
    3:36
    Collaborating to Accelerate COVID-19 Research
    Video
    Growth of Mouse Pancreatic Organoids in PancreaCult™ Culture Medium: Under a Microscope
    1:07
    Growth of Mouse Pancreatic Organoids in PancreaCult™ Culture Medium: Under a Microscope
    Video
    Growth of Liver Organoids in HepatiCult™ Culture Medium: Under a Microscope
    1:18
    Growth of Liver Organoids in HepatiCult™ Culture Medium: Under a Microscope
    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
    Spotlight on Organoids: Expert Panel
    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
    Patient-Derived Organoids for Drug Screening and Development
    50:25
    Patient-Derived Organoids for Drug Screening and Development
    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
    Modeling Host-Microbe Interactions Using Human Intestinal Organoids
    16:31
    Modeling Host-Microbe Interactions Using Human Intestinal Organoids
    Webinar
    ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases, Nephrotoxicity, and Highly Relevant Cell-Based Assay
    59:52
    ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases, Nephrotoxicity, and Highly Relevant Cell-Based Assay
    Webinar
    Adding Hepatic Organoids to Your Research
    50:59
    Adding Hepatic Organoids to Your 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
    Organoid Expert Panel
    52:35
    Organoid Expert Panel
    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
    Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
    56:31
    Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
    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
    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

    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. 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.
    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.
    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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