IntestiCult™-SF Organoid Growth Medium (Human)

Serum-free and conditioned medium-free cell culture medium for establishment and maintenance of human intestinal organoids

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IntestiCult™-SF Organoid Growth Medium (Human)

Serum-free and conditioned medium-free cell culture medium for establishment and maintenance of human intestinal organoids

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Serum-free and conditioned medium-free cell culture medium for establishment and maintenance of human intestinal organoids

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Product 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

Serum- and conditioned medium-free formulation

Suitable for use in genome editing protocols

Enables generation of intestinal organoids in one week

Complete medium formulation that delivers consistent results

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

Gentle Cell Dissociation Reagent
cGMP, enzyme-free cell dissociation reagent
Organoid Culture Plates
Cell culture plates for easy and reproducible generation of organoids
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
Y-27632 (Dihydrochloride)
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
Labeling Antibodies
Compatible antibodies for purity assessment of isolated cells

Overview

IntestiCult™-SF Organoid Growth Medium (Human) is a complete cell culture medium for efficient establishment and long-term maintenance of organoids derived from human intestinal crypts. IntestiCult™-SF is a serum-free and conditioned medium-free formulation.

Intestinal organoids provide a convenient in vitro organotypic culture system for studying the intestinal epithelium. Isolated intestinal crypts rapidly form complex organoids when cultured in IntestiCult™-SF. The organoids incorporate a functional lumen enclosed by a polarized epithelial cell layer containing known cell types of the adult intestinal epithelium. Maturing intestinal organoids establish protruding buds, resembling intestinal crypts of the in vivo tissues. IntestiCult™-SF supports the proliferation of intestinal epithelial stem cells from patient’s biopsies and expansion of organoid cultures derived from fresh tissue or from already established frozen stocks.

Applications of intestinal organoid cultures include studying the development and function of intestinal epithelium, modeling intestinal diseases, and performing targeted molecule screens. Intestinal organoid cultures can also be used for investigation of adult stem cell properties and for regenerative therapy approaches, and can be further differentiated using IntestiCult™ Organoid Differentiation Medium (Human)(Catalog #100-0214).

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™. Additionally, download our detailed e-book Proven Protocols for Intestinal Organoid Culture: Getting Started with IntestiCult™ for a curated collection of intestinal organoid protocols.

Should you intend to use this product for commercial purposes, please contact HUB Organoids B.V. at for a commercial use license or for clarifications in relation to HUB Organoids B.V. licensing.

Specialized Media

Intestinal Cells

Human

Organoid Culture

IntestiCult

Disease Modeling, Epithelial Cell Biology, Organoids, Stem Cell Biology

Serum-Free

Data Figures

Figure 1. Organoids Grown in IntestiCult™-SF Have a Comparable Morphology to Organoids Grown in IntestiCult Organoid Growth Medium (Human)

Human intestinal organoids grown in IntestiCult™-SF and IntestiCult™ Organoid Growth Medium (Human) display similar morphological characteristics during organoid maintenance and expansion. Shown are organoids from small intestinal and colonic tissues, established and expanded in IntestiCult™-SF or IntestiCult™ OGMH. Organoids are imaged at passage 5 of culture for all images. Scale bars = 500 μm.

Figure 2. IntestiCult™-SF Provides More Efficient Expansion of Intestinal Organoids Compared to IntestiCult™ OGMH

Human intestinal organoids established from both (A) small intestinal and (B) colonic tissues display more efficient expansion during extended culture when grown in IntestiCult™-SF compared to those grown in IntestiCult™ OGMH. Shown is the cumulative expansion of organoids averaged across two separate donor lines in each medium. IntestiCult™-SF demonstrates more efficient expansion of both small intestinal and colonic organoids during extended culture.

Figure 3. Organoids Grown in IntestiCult™-SF Display Some Characteristics of the Mature Intestinal Epithelium

While organoids grown in IntestiCult™-SF display a primarily "immature" phenotype including expression of (A) intestinal stem cell marker OLFM4 and (B) proliferation marker ki67. (C, D) These organoids also display some characteristics of the mature epithelium such as the presence of goblet cells (C, Muc2), enterocytes (C, KRT20), and enteroendocrine cells (D, CHGA). Scale bars = 50 μm or 20 μm as indicated.

Figure 4. Organoids Grown in IntestiCult™-SF Display Similar Genetic Expression Profiles to Those Grown in IntestiCult™ OGMH

Analysis of intestinal organoids grown in IntestiCult™-SF by qPCR of (A) Lgr5, (B) Muc2, (C) Krt20, and (D) ChgA demonstrates similar genetic expression profiles to those grown in IntestiCult™ OGMH, as well as to colonic and small intestinal tissue (A-D; x and + respectively). All expression levels are shown relative to ACTB and TBP house-keeping genes (HKGs). (E) Further analysis of three separate donors grown in IntestiCult™-SF (circles) IntestiCult™ OGMH (triangles), and published medium (squares) by principle component analysis demonstrates that differences between cultures are primarily due to donor variability with samples forming distinct clusters separated by donor, rather than by medium.

Figure 5. IntestiCult™-SF Enables Efficient Expansion of Intestinal Organoids from Polyp Tissue

Intestinal organoids were established from both normal epithelial tissue, as well as from polyp tissue, and expanded for three passages in IntestiCult™-SF. Shown are cultures from two separate donors, imaged at the end of each passage immediately before passaging.

Figure 6. Organoids Grown in IntestiCult™-SF can be Further Differentiated using IntestiCult™ Organoid Differentiation Medium

Further differentiation of organoids in IntestiCult™-SF can be achieved by passaging organoid cultures in IntestiCult™ Organoid Differentiation Medium as organoid-derived monolayers (2D Monolayer Diff) or in 3D organoid culture (3D Organoid Diff). Upon differentiation, markers for enterocytes (KRT20, ApoB), goblet cells (Muc2), and enteroendocrine cells (ChgA) are upregulated compared to organoids grown in IntestiCult™-SF.

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

Document Type

Product Name

Catalog #

100-0340

Lot #

All

Language

English

Document Type

Product Name

Catalog #

100-0340

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.

Research Area

Workflow Stages

Workflow Stages for Intestinal Epithelial Cell Research

Tissue Dissociation

Cell Culture

Workflow Stages for Organoids

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Resources and Publications

An integrated patient-derived colon organoids platform as a functional model for nutraceutical and stress response. A. Costantino et al. iScience 2026 Jun

Abstract

Nutraceuticals are increasingly investigated for their capacity to modulate oxidative and inflammatory stress, yet preclinical testing still relies largely on immortalized cell lines or animal models that poorly recapitulate human epithelial complexity. To address this gap, we developed an integrated platform based on patient-derived colon organoids generated from non-tumoral mucosa and maintained under proliferative or differentiation conditions to model distinct epithelial states. The system combines millifluidic measurement of individual organoid mass, density, and diameter with bulk RNA sequencing and digital PCR profiling to enable multiparametric characterization. Transcriptional analysis revealed state-specific gene programs and shifts in epithelial and immune-related pathways, while biophysical measurements captured structural remodeling. In this pilot validation, a defined oxidative insult followed by nutraceutical treatment elicited coordinated transcriptional and phenotypic responses. This integrated approach provides a scalable and physiologically relevant framework for functional nutraceutical profiling and mechanistic studies of epithelial stress responses.

Tailoring a novel colorectal cancer stem cell-targeted therapy by inhibiting the SMYD3/c-MYC axis M. L. Signorile et al. Signal Transduction and Targeted Therapy 2025 Jun

Abstract

Cancer stem cells (CSCs) are responsible for colorectal cancer (CRC) chemoresistance, recurrence, and metastasis. Therefore, identifying molecular stemness targets that are involved in tumor growth is crucial for effective treatment. Here, we performed an extensive in vitro and in vivo molecular and functional characterization, revealing the pivotal role of the lysine methyltransferase SET and MYND Domain Containing 3 (SMYD3) in colorectal cancer stem cell (CRC-SC) biology. Specifically, we showed that SMYD3 interacts with and methylates c-MYC at K158 and K163, thereby modulating its transcriptional activity, which is implicated in stemness and colorectal malignancy. Our in vitro data suggest that SMYD3 pharmacological inhibition or its stable genetic ablation affects the clonogenic and self-renewal potential of patient-derived CRC-SCs and organoids by altering their molecular signature. Moreover, we found that SMYD3 stable knock-out or pharmacological inhibition drastically reduces CRC tumorigenicity in vivo and CRC-SC metastatic potential. Overall, our findings identify SMYD3 as a promising therapeutic target acting directly on c-MYC, with potential implications for countering CRC-SC proliferation and metastatic dissemination. Subject terms: Gastrointestinal cancer, Cancer stem cells

Robust and reproducible human intestinal organoid-derived monolayer model for analyzing drug absorption K. Tanaka et al. Scientific Reports 2025 Apr

Abstract

Predicting the absorption of orally administered drugs is crucial to drug development. Current in vitro models lack physiological relevance, robustness, and reproducibility, thus hindering reliable predictions. In this study, we developed a reproducible and robust culture method to generate a human intestinal organoid-derived monolayer model that can be applied to study drug absorption through a step-by-step approach. Our model showed similarity to primary enterocytes in terms of the drug absorption-related gene expression profile, tight barrier function, tolerability toward artificial bile juice, drug transporter and metabolizing enzyme function, and nuclear receptor activity. This method can be applied to organoids derived from multiple donors. The permeability of launched 19 drugs in our model demonstrated a correlation with human Fa values, with an R 2 value of 0.88. Additionally, by combining the modeling and simulation approaches, the estimated FaFg values for seven out of nine drugs, including CYP3A substrates, fell within 1.5 times the range of the human FaFg values. Applying this method to the drug discovery process might bridge the gap between preclinical and clinical research and increase the success rates of drug development.

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Interested in trying ��ƽ��’s organoid products for your intestinal research? Fill out the form to request information about introductory offers.

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 it for drug screening aiming for commercial gain, equipment validation, biobanking, or for other commercial purposes. Purchasers wishing to use the product for purposes other than basic research use should contact HUB at www.huborganoids.nl to obtain a further license. Purchasers may apply for a License from HUB, which 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 ��ƽ��, REFER TO WWW.��ƽ��.COM/COMPLIANCE.

Safety Statement:

CA WARNING: This product can expose you to Progesterone which is known to the State of California to cause cancer. For more information go to

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IntestiCult™-SF Organoid Growth Medium (Human)

Serum-free and conditioned medium-free cell culture medium for establishment and maintenance of human intestinal organoids

IntestiCult™-SF Organoid Growth Medium (Human)

Serum-free and conditioned medium-free cell culture medium for establishment and maintenance of human intestinal organoids

Product Advantages

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Overview

Data Figures

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (46)

Publications (5)

Abstract

Abstract

Abstract

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