PancreaCult™ Organoid Growth Medium (Mouse)

Cell culture medium for establishment and maintenance of mouse pancreatic exocrine organoids
PancreaCult™ Organoid Growth Medium (Mouse)

Cell culture medium for establishment and maintenance of mouse pancreatic exocrine organoids

1 Kit
Catalog # 06040
299 USD
Required Products
  1. AggreWell™ Rinsing Solution
    Anti-Adherence Rinsing Solution

    Rinsing solution for cultureware to prevent cell adhesion

  2. Dispase (1 U/mL)
    Dispase (1 U/mL)

    1 U/mL dispase in DMEM/F-12

  3. Collagenase Type IV (1 mg/mL)
    Collagenase Type IV (1 mg/mL)

    Cell dissociation reagent

Overview

PancreaCult™ Organoid Growth Medium (Mouse) is a serum-free, defined cell culture medium for establishment and maintenance of mouse pancreatic exocrine organoids. These organoids, or "mini-pancreata", provide an in vitro organotypic culture system for studying pancreatic cell biology, disease, and cancer. Organoids grown in PancreaCult™ feature an epithelium expressing genes marking pancreatic stem cells (LGR5), progenitor cells (PDX1, SOX9), and ductal cells (CAR2, MUC1, KRT19). Pancreatic organoids can be passaged every 3 - 6 days for long-term maintenance and can also be cryopreserved.

PancreaCult™ supports mouse pancreatic organoid culture either embedded in Corning® Matrigel® domes or in a dilute Matrigel® suspension. Organoid culture enables convenient in vitro characterization of the pancreatic epithelium in a physiologically relevant system and reduces the need for animal use.
Advantages
• Convenient in vitro system for generating organoids within one week
• Detailed, 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
• PancreaCult™ OGM Mouse Basal Medium, 95 mL
• PancreaCult™ OGM Mouse Supplement, 5 mL
Subtype
Specialized Media
Cell Type
Pancreatic Cells
Application
Cell Culture, Expansion, Maintenance, Organoid Culture
Brand
PancreaCult
Area of Interest
Cancer Research, Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Stem Cell Biology, Transplantation Research
Formulation
Serum-Free

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
PancreaCult™ Organoid Growth Medium (Mouse)
Catalog #
06040
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
PancreaCult™ Organoid Growth Medium (Mouse)
Catalog #
06040
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
PancreaCult™ Organoid Growth Medium (Mouse)
Catalog #
06040
Lot #
All
Language
English

Educational Materials(14)

Brochure
2019-2020 Cell Culture Training Catalog
Brochure
PancreaCult™ Organoid Growth Medium (Mouse)
Brochure
Organoids
Technical Bulletin
Mouse Pancreatic Exocrine Organoid Culture: Supplementary Protocols
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
Webinar
1:10:09
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
Webinar
Spotlight on Organoids: Expert Panel
Webinar
41:47
Challenges in Translating iPSC Technology
Webinar
50:59
Adding Hepatic Organoids to Your Research
Webinar
24:56
GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
Webinar
52:35
Organoid Expert Panel
Scientific Poster
Culture and Differentiation of Mouse Hepatic Organoids Using Serum-free HepatiCult™ Organoid Growth Medium
Scientific Poster
Modelling Pancreatic Cancer Through Pancreatic Exocrine Organoids Using PancreaCult™ Serum-free Medium
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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

Figure 1. Organoids Grown in PancreaCult™ Organoid Growth Medium (Mouse)

Pancreatic exocrine organoids are observed within one week when cultured in (A) Corning® Matrigel® domes or (B) a dilute Matrigel® suspension. Organoids were imaged during passage 2, on day 4.

Figure 2. Mouse Pancreatic Organoids can be Initiated from a Variety of Starting Materials

PancreaCult™ Organoid Growth Medium (Mouse) enables the initiation of pancreatic exocrine organoids from (A) duct fragments, (B) single cells and (C) cryopreserved organoid fragments. All organoids were grown in Matrigel® domes. Organoids were imaged on day 4 or day 5 of primary culture (duct fragments and single cells, respectively) or day 3 of the first passage post-thaw (cryopreserved organoids).

Figure 3. Pancreatic Organoids can be Grown in Matrigel® Domes or as a Dilute Matrigel® Suspension

Organoids cultured using PancreaCult™ Organoid Growth Medium (Mouse) from freshly isolated pancreatic tissue fragments and plated in (A) Matrigel® domes or (B) as a dilute Matrigel® suspension. Organoids grown in either culture condition are typically ready for passage within 3 - 6 days.

Figure 4. Pancreatic Exocrine Organoids Display Markers of Pancreatic Progenitor and Ductal Cells

Pancreatic exocrine organoids grown in PancreaCult™ and stained for nuclei (DAPI, blue), ductal marker KRT19 (green) and pancreatic progenitor marker PDX1 (red). Organoids were imaged during passage 12 on day 5. Note: The folded appearance of epithelium is a function of cryosectioning and not representative of the shape of proliferating organoids.

Figure 5. Pancreatic Exocrine Organoids Retain Pancreatic Marker Expression During Passaging

Pancreatic organoids express stem cell markers and those typical of the pancreatic exocrine system, including (A) Axin2, (B) Krt19, (C) Muc1 and (D) Pdx1. Relative quantification (RQ) of each marker is reported relative to the 18S and TBP housekeeping genes and normalized to C57/Bl6 pancreatic tissue. Marker expression was assayed during early passages (passage 1-5) and late passages (passage 6-10).

Figure 6. Expansion of Organoids Grown in PancreaCult™ Organoid Growth Medium (Mouse)

Organoids cultured with PancreaCult™ Organoid Growth Medium (Mouse) show efficient growth over multiple passages. Cultures were split with an average split ratio of 1:16 at each passage.

Figure 7. Pancreatic Exocrine Organoids Provide a Model for Pancreatic Carcinomas

PancreaCult™ Organoid Growth Medium (Mouse) supports the growth of organoids from pancreatic carcinomas. Pancreatic ducts were isolated from KPC mice (Kras+/LSL-G12D; Trp53+/LSL-R172H; Pdx1-Cre) and cultured in PancreaCult™ Organoid Growth Medium (Mouse). Organoids were imaged on (A) day 4 of primary culture and (B) day three after the first passage. An activated KRAS genotype was retained in organoids during culture. Data used with permission from Dr. David Tuveson.

Publications (1)

Cell 2019 A Multiscale Map of the Stem Cell State in Pancreatic Adenocarcinoma. N. K. Lytle et al.

Abstract

Drug resistance and relapse remain key challenges in pancreatic cancer. Here, we have used RNA sequencing (RNA-seq), chromatin immunoprecipitation (ChIP)-seq, and genome-wide CRISPR analysis to map the molecular dependencies of pancreatic cancer stem cells, highly therapy-resistant cells that preferentially drive tumorigenesis and progression. This integrated genomic approach revealed an unexpected utilization of immuno-regulatory signals by pancreatic cancer epithelial cells. In particular, the nuclear hormone receptor retinoic-acid-receptor-related orphan receptor gamma (ROR$\gamma$), known to drive inflammation and T cell differentiation, was upregulated during pancreatic cancer progression, and its genetic or pharmacologic inhibition led to a striking defect in pancreatic cancer growth and a marked improvement in survival. Further, a large-scale retrospective analysis in patients revealed that ROR$\gamma$ expression may predict pancreatic cancer aggressiveness, as it positively correlated with advanced disease and metastasis. Collectively, these data identify an orthogonal co-option of immuno-regulatory signals by pancreatic cancer stem cells, suggesting that autoimmune drugs should be evaluated as novel treatment strategies for pancreatic cancer patients.
View All Publications

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