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HepatiCult™ Organoid Growth Medium (Mouse)

Cell culture medium for establishment and maintenance of mouse hepatic progenitor organoids

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HepatiCult™ Organoid Growth Medium (Mouse)

Cell culture medium for establishment and maintenance of mouse hepatic progenitor organoids

1 Kit
Catalog #06030
264 USD

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Overview

HepatiCult™ Organoid Growth Medium (Mouse) is a serum-free, defined cell culture medium for establishment and maintenance of mouse hepatic progenitor organoids. These organoids, or "mini-livers", provide an in vitro organotypic culture system for studying hepatic stem and progenitor cells. Organoids grown in HepatiCult™ feature an epithelium expressing genes marking hepatic stem and progenitor cells (PROM1, AXIN2, SOX9 and CD44), ducts (KRT19 and HNF1b) and hepatocytes (HNF4a, AFP). Hepatic organoids can be passaged every 4 - 7 days, can be cryopreserved, and are primed for downstream differentiation.

HepatiCult™ supports mouse hepatic organoid culture either embedded in Corning® Matrigel® domes or in a dilute Matrigel® suspension. Organoid culture enables convenient in vitro characterization of the hepatic epithelium in a physiologically relevant system and reduces the need for animal use.
Advantages:
• Convenient in vitro system for generating organoids in 4 - 5 days
• Step-by-step protocol with no injury models, hand-picking of ducts, or cell sorting required
• Simple, two-component format; serum-free and defined medium formulation
• Flexible protocol for generation and maintenance of organoid cultures in matrix domes or suspension culture
Components:
• HepatiCult™ OGM Mouse Basal Medium, 95 mL
• HepatiCult™ OGM Mouse Supplement, 5 mL
Subtype:
Specialized Media
Cell Type:
Hepatic Cells
Species:
Mouse
Application:
Cell Culture; Expansion; Maintenance; Organoid Culture
Brand:
HepatiCult
Area of Interest:
Cancer Research; Disease Modeling; Drug Discovery and Toxicity Testing; Epithelial Cell Biology; Stem Cell Biology
Formulation:
Serum-Free; Defined

Scientific Resources

Educational Materials

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Data and Publications

Data

Figure 1. Mouse Hepatic Progenitor Organoids can be Initiated from a Variety of Starting Materials

HepatiCult™ Organoid Growth Medium (Mouse) enables the initiation of hepatic progenitor organoids from (A) duct fragments, (B) single cells or (C) cryopreserved organoids. All organoids were grown in Matrigel® domes and imaged on day 7 of primary culture or the first passage post thaw (cryopreserved organoids).

Figure 2. Hepatic Organoids can be Grown in Matrigel® Domes or as a Dilute Matrigel® Suspension

Hepatic progenitor organoids were cultured from freshly isolated mouse hepatic duct fragments in HepatiCult™ Organoid Growth Medium (Mouse) and plated (A) in Matrigel® domes or (B) as a dilute Matrigel® suspension. Organoids grown in either culture condition are ready for passage within 4-7 days.

Figure 3. Organoids grown in HepatiCult™ Organoid Growth Medium (Mouse) Display Some Characteristics Typical of the Mature Hepatic Epithelium

(A) Hepatic progenitor organoids exhibit the polygonal morphology typical of the hepatic epithelium. (B) Hepatic progenitors cultured as organoids show binucleation (arrows), a feature common of mature hepatocytes. (C) Immunocytochemistry analysis shows localization of MRP4 (green), a membrane-bound, unidirectional efflux transporter, along the exterior of the organoids and DAPI (red) localized to the cellular nuclei. This indicates cellular polarization of the organoids with the basolateral surface of the epithelium distal from the lumen. (D) Hepatic organoids contain an actively dividing progenitor population, seen through the expression of Ki67 (red). Cell nuclei are stained with DAPI (blue).

Figure 4. Analysis of Organoid Gene Expression when Grown in Matrigel® Domes and as a Dilute Matrigel® Suspension

Analysis of marker expression by RNA-seq shows organoids grown in HepatiCult™ Organoid Growth Medium (Mouse) in either Matrigel® domes or in a dilute Matrigel® suspension express markers associated with hepatic stem and progenitor cells. The organoids also express low levels of genes associated with mature hepatic cell types, including cholangiocytes and hepatocytes. Columns represent biological replicates at passages ranging from passage 1 - 40.

Figure 5. Expansion of Organoids grown in HepatiCult™ Organoid Growth Medium (Mouse)

Organoids cultured with HepatiCult™ Organoid Growth Medium (Mouse) show efficient growth over passaging. Cultures were split with an average split ratio of 1:25 at each passage.

Figure 6. Differentiation of Hepatic Progenitor Organoids

Hepatic progenitor organoids grown in HepatiCult™ Organoid Growth Medium (Mouse) (EM) can be differentiated to resemble more mature cell types when switched to a differentiation medium (DM)1,2. After switching to published differentiation medium hepatic organoids show the upregulation of mature hepatic markers such as (A) Hnf4α and (B) Alb, downregulation of hepatic stem cell and progenitor markers (C) Sox9 and (D) Axin2 and upregulation of ductal markers (E) Krt19 and (F) Hnf1β. Relative quantification (RQ) for each marker is reported relative to 18S and TBP housekeeping genes and normalized relative to hepatic progenitor organoids grown in HepatiCult™ Organoid Growth Medium (Mouse) and cultured in Matrigel® domes.

STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.