HepatiCult™ Organoid Kit (Human)

Culture medium kit for initiation, growth, and differentiation of human liver organoids
HepatiCult™ Organoid Kit (Human)

Culture medium kit for initiation, growth, and differentiation of human liver organoids

1 Kit
Catalog # 100-0386
995 USD

INDIVIDUAL COMPONENTS

HepatiCult™ Organoid Initiation Medium (Human)

479 USD
Catalog # 100-0384
1 Kit

Culture medium to initiate organoids from primary liver tissue

HepatiCult™ Organoid Growth Medium (Human)

349 USD
Catalog # 100-0385
1 Kit

Culture medium for long-term expansion and maintainence of human liver organoids

HepatiCult™ Organoid Differentiation Medium (Human)

239 USD
Catalog # 100-0383
1 Kit

Culture medium kit for differentiation of human liver organoids

Required Products
  1. Trypan Blue
    Trypan Blue

    Reagent for counting viable mammalian cells

  2. Ammonium Chloride Solution|07800
    Ammonium Chloride Solution

    Reagent for lysis of red blood cells

  3. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

  4. Costar® 24-Well Flat-Bottom Plate, Tissue Culture-Treated
    Costar® 24-Well Flat-Bottom Plate, Tissue Culture-Treated

    Clear polystyrene flat-bottom, tissue culture-treated multiwell cell culture plate with lid

Overview

Reliably generate, expand, and differentiate physiologically relevant human liver organoids. Compared to traditionally used model systems such as cell lines and animal models, human liver organoids provide a more relevant model system for studying hepatic development, regeneration, metabolism, and disease. The HepatiCult™ Organoid Kit (Human) (Catalog #100-0386) contains all the necessary components to culture human liver organoids from fresh or cryopreserved human liver tissue. HepatiCult™ Organoid Initiation Medium (Catalog #100-0384) efficiently generates hepatic organoids across different donor lines, which can be further expanded and maintained in HepatiCult™ Organoid Growth Medium (Catalog #100-0385) for future experimentation or bio-banking. Differentiating these organoids with HepatiCult™ Organoid Differentiation Medium (Catalog #100-0383) generates mature organoids that demonstrate hepatic functionality, including CYP3A4 activity. These organoids can be adapted to a range of culture protocols, including 2D monolayer, suspension cultures, and high-throughput assays.

Learn more about the supplementary protocols.
Advantages
⦁ Provides a complete culture system for establishing, maintaining, and differentiating human liver organoids
⦁ Enables efficient organoid initiation across different donor liver tissues
⦁ Generates mature hepatic organoids that demonstrate CYP3A4 activity
⦁ Allows experimental flexibility with different culture formats
Components
  • HepatiCult™ Organoid Kit (Human), Catalog #100-0386
    • HepatiCult™ Organoid Basal Medium (Human), 3 x 95 mL
    • Organoid Supplement, 50 mL
    • HepatiCult™ Organoid Growth Supplement (Human), 2 x 5 mL
    • HepatiCult™ Organoid Differentiation Supplement (Human), 5 mL
  • HepatiCult™ Organoid Initiation Medium (Human), Catalog #100-0384
    • HepatiCult™ Organoid Basal Medium (Human), 95 mL
    • Organoid Supplement, 50 mL
    • HepatiCult™ Organoid Growth Supplement (Human), 5 mL
  • HepatiCult™ Organoid Growth Medium (Human), Catalog #100-0385
    • HepatiCult™ Organoid Basal Medium (Human), 95 mL
    • HepatiCult™ Organoid Growth Supplement (Human), 5 mL
  • HepatiCult™ Organoid Differentiation Medium (Human), Catalog #100-0383
    • HepatiCult™ Organoid Basal Medium (Human), 95 mL
    • HepatiCult™ Differentiation Supplement (Human), 5 mL
Subtype
Specialized Media
Cell Type
Hepatic Cells
Species
Human
Application
Organoid Culture
Brand
HepatiCult
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Organoids, Stem Cell Biology
Formulation
Serum-Free

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
HepatiCult™ Organoid Kit (Human)
Catalog #
100-0386
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
HepatiCult™ Organoid Initiation Medium (Human)
Catalog #
100-0384
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
HepatiCult™ Organoid Growth Medium (Human)
Catalog #
100-0385
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
HepatiCult™ Organoid Differentiation Medium (Human)
Catalog #
100-0383
Lot #
All
Language
English
Document Type
Manual
Product Name
HepatiCult™ Organoid Kit (Human)
Catalog #
100-0386
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
HepatiCult™ Organoid Kit (Human)
Catalog #
100-0386
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
HepatiCult™ Organoid Kit (Human)
Catalog #
100-0386
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
HepatiCult™ Organoid Kit (Human)
Catalog #
100-0386
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
HepatiCult™ Organoid Kit (Human)
Catalog #
100-0386
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
HepatiCult™ Organoid Initiation Medium (Human)
Catalog #
100-0384
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
HepatiCult™ Organoid Initiation Medium (Human)
Catalog #
100-0384
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
HepatiCult™ Organoid Initiation Medium (Human)
Catalog #
100-0384
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
HepatiCult™ Organoid Growth Medium (Human)
Catalog #
100-0385
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
HepatiCult™ Organoid Growth Medium (Human)
Catalog #
100-0385
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
HepatiCult™ Organoid Differentiation Medium (Human)
Catalog #
100-0383
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
HepatiCult™ Organoid Differentiation Medium (Human)
Catalog #
100-0383
Lot #
All
Language
English

Educational Materials (8)

Protocol
Establishment of 3D Organoid-Derived Hepatic Monolayers Using HepatiCult™ Organoid Initiation Medium (Human)
Protocol
Recovery and Expansion of Hepatic Organoids Using HepatiCult™ Organoid Growth Medium
Protocol
Performing Immunocytochemical Staining of Epithelial Organoids
Protocol
Expansion of Hepatic Organoids in Dilute Matrigel® Suspension Cultures Using HepatiCult™ Organoid Growth Medium
Protocol
Cryopreserving Human Liver Tissue for Organoid Culture
Protocol
Expansion of Hepatic Organoids via Single Cells Using HepatiCult™ Organoid Growth Medium
Protocol
Cryopreserving Hepatic Organoids Expanded in HepatiCult™ Organoid Growth Medium (Human)
Webinar
Hepatic Organoids: Improved Tools and In Vitro Models for Drug Development
16:13
Hepatic Organoids: Improved Tools and In Vitro Models for Drug Development

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 Organoids

Data and Publications

Data

HepatiCult™ Organoid Kit (Human) Enables Liver Organoid Initiation, Expansion, and Differentiation

Figure 1. HepatiCult™ Organoid Kit (Human) Enables Liver Organoid Initiation, Expansion, and Differentiation

Human liver organoids can be grown from normal human liver tissue-derived hepatic ducts using the HepatiCult™ Organoid Kit (Human). (A) Cultures are established in HepatiCult™ Organoid Initiation Medium (OIM; Human) or HepatiCult™ Organoid Growth Medium (OGM; Human) (see Table 1 below) and subsequently passaged in HepatiCult™ OGM for expansion. (B) After passaging 2-3 times in HepatiCult™ OGM, cultures can be switched to HepatiCult™ Organoid Differentiation Medium (ODM; Human) to differentiate organoids towards more mature hepatic cell types. Refer to the Product Manual (Document #10000008301) for full culturing protocols.

Table 1. Product Recommendations for Liver Organoid Initiation, Expansion, and Differentiation

The recommended configuration of the HepatiCult™ Organoid Kit (Human) may differ based on starting material and experimental goals. When establishing liver organoid cultures from human liver tissue, HepatiCult™ Organoid Initiation Medium (OIM; Human) is recommended for efficient organoid initiation (see Figure 2 below). The expansion of already established organoids (fresh in culture or cryopreserved) is supported by HepatiCult™ Organoid Growth Medium (OGM; Human). These organoids should be maintained for 2-3 passages before further differentiation using HepatiCult™ Organoid Differentiation Medium (ODM). Refer to the Product Manual (Document #10000008301) for full culturing protocols.

HepatiCult™ Organoid Kit (Human) Provides Efficient Organoid Initiation From Human Liver Tissue

Figure 2. HepatiCult™ Organoid Kit (Human) Provides Efficient Organoid Initiation From Human Liver Tissue

(A) Organoid cultures were initiated in HepatiCult™ OIM, and then (B) passaged into HepatiCult™ OGM. For serum-free culture conditions, organoid cultures were both, (C) initiated in and (D) passaged in HepatiCult™ OGM. Culture images shown are from (A, C) day 15 following initiation, and (B, D) on day 8 of the first passage. (E) Quantification of organoid initiation efficiency shows a significantly higher organoid yield in HepatiCult™ OIM per gram of human liver tissue (mean ± SD; n=14).

HepatiCult™ Organoid Initiation Medium (Human) Supports Robust Organoid Establishment

Figure 3. HepatiCult™ Organoid Initiation Medium (Human) Supports Robust Organoid Establishment Across Multiple Liver Tissue Donor Samples

Organoids initiated from 4 donor tissue samples (A-D) exhibit morphological heterogeneity 15 days after initiation. All initiated cultures were subsequently expanded in HepatiCult™ OGM using a 1:1 passaging ratio (E-H), yielding healthy organoids at the end of the first passage.

Expansion of Organoid Cultures in HepatiCult™ Organoid Growth Medium

Figure 4. Expansion of Organoid Cultures in HepatiCult™ Organoid Growth Medium

Hepatic organoids show efficient growth in HepatiCult™ OGM across multiple donors and passages with potential for indefinite culture. Organoids initiated using the serum-free workflow exhibited comparable growth rates when expanded in HepatiCult™ OGM (open markers).

Proliferating Hepatic Organoids Display Characteristics of Hepatic Progenitors

Figure 5. Proliferating Hepatic Organoids Display Characteristics of Hepatic Progenitors

Human liver organoids grown in HepatiCult™ OGM display characteristics of proliferating hepatic progenitors observed through immunocytochemistry staining of (A) KI67, (B) HNF4A and (C) SOX9. Proliferating hepatic organoids also display characteristics of the hepatic epithelium including expression of (A) EPCAM. (B, C) Nuclei are counterstained with DAPI.

Proliferating Hepatic Organoids Maintain Genetic Expression Across Multiple Passages

Figure 6. Proliferating Hepatic Organoids Maintain Genetic Expression Across Multiple Passages

Liver organoids maintained in HepatiCult™ OGM express stem cell markers (A) LGR5 and (B) AXIN2, ductal markers (C) SOX9 and (D) KRT19, as well as hepatic marker (E) HNF4a and (F) Albumin (ALB) across multiple passages, with minimal albumin expression observed during culture in HepatiCult™ OGM. Expression levels were measured by qPCR and normalized to TBP and UBC housekeeping genes to quantify relative expression levels. (mean ± SD; n = 2-5 organoid lines), * p < 0.05.

Organoid Differentiation Induces Changes in Organoid Morphology

Figure 7. Organoid Differentiation Induces Changes in Organoid Morphology

Organoids exhibit a compact and dense morphology, often comprising thickened epithelia, upon switching cultures to HepatiCult™ Organoid Differentiation Medium (ODM). Shown are images of the same culture well over the course of the differentiation, including (A) day two of culture in HepatiCult™ OGM, (B) day five of culture, immediately after switching organoid cultures from HepatiCult™ OGM to HepatiCult™ ODM, (C) day seven of culture (two days after switching to ODM), (D) day ten of culture (five days after switching to ODM), and (E) day 15 of culture (ten days after switching to ODM). (F) Magnification of the rectangular section highlighted in (E).

Differentiation of Hepatic Organoids in HepatiCult™ ODM Induces Changes in Gene Expression Consistent With Hepatic Maturation

Figure 8. Differentiation of Hepatic Organoids in HepatiCult™ ODM Induces Changes in Gene Expression Consistent With Hepatic Maturation

Upon differentiation in HepatiCult™ ODM, liver organoids show changes in gene expression with a decrease in expression of stem cell marker LGR5, as well as of ductal markers CK19, CK7, and SOX9, and an increase in expression of hepatic markers HNF4A, ALB, CYP3A4, and ASGR1. Each replicate represents an individual donor sample analyzed by qPCR at passage 4 on day 8 in HepatiCult™ OGM and day 15 in HepatiCult™ ODM, showing relative gene expression levels normalized to TBP and UBC housekeeping genes.

Differentiated Hepatic Organoids Demonstrate Functionality of Mature Hepatocytes

Figure 9. Differentiated Hepatic Organoids Demonstrate Functionality of Mature Hepatocytes

Upon differentiation in HepatiCult™ ODM, liver organoids were assayed for (A) albumin secretion, (B) CYP3A4 activity, (C) total bile acid production, and (D) urea production. Hepatic functionalities were compared to HepG2 cells and primary human hepatocytes (PHH), which were cultured in supplier-recommended media. Albumin secretion was detected using an ELISA kit (Abcam), total bile acid and urea production were analysed using colorimetric kits (Abcam), and CYP43A4 activity, referring to baseline activity without induction, was determined using the Luciferin-IPA kit (Promega). (mean ± SD; n = 3 organoid lines across 2 experiments, n = 2-3 technical replicates of HepG2 in 1 experiment, and n = 3 cryopreserved PHH donor samples in 1 experiment), * p < 0.05; ** p < 0.01.

HepatiCult™ Organoid Kit Supports the Growth and Differentiation of Porcine Liver Organoids

Figure 10. HepatiCult™ Organoid Kit Supports the Growth and Differentiation of Porcine Liver Organoids

Porcine liver organoids were established in HepatiCult™ Organoid Initiation Medium, and subsequently expanded for 4 passages in (A) HepatiCult™ Organoid Growth Medium (Human) and differentiated in (B, C) HepatiCult™ Organoid Differentiation Medium (Human). Organoids are stained with basolateral protein marker p120 (green), apical membrane and bile canaliculi marker F-Actin (purple), and nuclear dye Hoechst (blue). (D) Magnification of Panel C indicates bile canaliculi formation as seen through F-Actin staining. Scale bars = 50 μm. Data used with permission from Dr. Amy Engevik (Vanderbilt University Medical Center)

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