PancreaCult™ Organoid Media (Human)

Culture media kits for initiation, growth, and establishment of human pancreatic organoids
概要

PancreaCult™ (Human) culture medium system provides a complete and robust workflow to establish, expand, and maintain pancreatic duct organoids. Pancreatic organoids are established from fresh or cryopreserved islet-depleted exocrine fractions in PancreaCult™ Organoid Initiation Medium (OIM), which is optimized to provide higher rates of organoid formation from tissue. These organoids are then transferred to PancreaCult™ Organoid Growth Medium (OGM) for long-term maintenance and passaging of pancreatic organoids, or for cryopreservation for future experiments. Organoids grown using the PancreaCult™ (Human) medium system can be adapted to various culture protocols, including two-dimensional Transwell® monolayer cultures, dilute Corning® Matrigel® suspension cultures, and highthroughput assay-compatible culture formats.

Advantages
• Convenient, in vitro system that recapitulates many key characteristics of adult pancreatic ducts
• Enables efficient organoid initiation and expansion across different pancreatic donor tissues
• Flexible growth medium allow expansion of normal and tumor-derived organoids
• Serum-free and conditioned medium-free
Components
  • PancreaCult™ Organoid Initiation Medium (Human) (Catalog #100-0820)
    • PancreaCult™ Organoid Basal Medium (Human) , 95 mL
    • PancreaCult™ Organoid Growth Supplement (Human), 5 mL
    • Organoid Supplement, 50 mL
  • PancreaCult™ Organoid Growth Medium (Human) (Catalog #100-0781)
    • PancreaCult™ Organoid Basal Medium (Human) , 95 mL
    • PancreaCult™ Organoid Growth Supplement (Human), 5 mL
Subtype
专业培养基
Cell Type
胰腺细胞
Species
Application
扩增, 类器官培养, 细胞培养, 维持培养
Brand
PancreaCult
Area of Interest
Organoids, 上皮细胞生物学, 干细胞生物学, 药物发现和毒性测试
Formulation
无血清


数据及文献

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

数据

Human Pancreatic Duct Organoids Grown using PancreaCult™ (Human)

Figure 1. Human Pancreatic Duct Organoids

PancreaCult™ Organoid Initiation Medium (OIM; Human) and Organoid Growth Medium (OGM; Human) support the initiation and expansion of human pancreatic duct organoids from pancreatic tissue or previously established organoid cultures. Shown are organoids grown using PancreaCult™ OIM and OGM and imaged on day 7 of passage 3.

Workflow for Generating Pancreatic Ductal Organoids Using PancreaCult™ (Human)

Figure 2. PancreaCult™ Human Enables Initiation and Expansion of Pancreatic Ductal Organoids

Human pancreatic ductal organoids are initiated in PancreaCult™ OIM for the first 3 days of culture, then switched to PancreaCult™ OGM for the remainder of culture. Alternatively, organoids can be grown in PancreaCult™ OGM for a completely serum-free protocol, however, proliferative pancreatic ductal cells are better supported in PancreaCult™ OIM during the first 3 days of culture. Cultures should be passaged after 7 days with further medium changes every 2-3 days. Pancreatic ductal organoids are suitable for experimentation or banking after one full passage in PancreaCult™ OGM. For full culture instructions please refer to the product manual (Document #10000011617)

PancreaCult™ (Human) Provides Robust Expansion of Pancreatic Duct Organoids

Figure 3. PancreaCult™ (Human) Provides Robust Expansion of Pancreatic Duct Organoids

(A) Organoid expansion in PancreaCult™ OGM provides robust expansion of organoids across different donors. (B) Organoids may be initiated in PancreaCult™ OGM for a completely serum-free workflow, however, initiation and maintenance of damaged tissue is better supported by initiating cultures in PancreaCult™ OIM. (C) Comparison of PancreaCult™ Human to two different DIY formulations showed more robust expansion of organoids in PancreaCult™ Human. Shown is the cumulative fold expansion of organoid fragments as counted at the end of each passage.

Pancreatic Duct Organoids Display Features of the Pancreatic Ductal Epithelium by ICC Staining

Figure 4. Pancreatic Duct Organoids Display Features of the Pancreatic Ductal Epithelium

Organoids grown using the PancreaCult™ (Human) display marker expression consistent with the pancreatic ductal epithelium when imaged using immunocytochemistry. Shown are organoids grown in PancreaCult™ OGM and stained for (A) pancreatic ductal marker CK19, (B) pancreatic ductal marker SOX9, (C) epithelial marker EPCAM, (D) proliferation marker KI67, (E) apical pancreatic duct marker MUC1, and (F) pancreatic ductal marker CA2. Organoids were imaged on passage 2 (A), passage 3 (B, C) or passage 10 (D-F).

Pancreatic Duct Organoids Cultured With PancreaCult™ Human Show Pancreatic Marker Expression Levels Similar to Exocrine Tissue

Figure 5. Pancreatic Duct Organoids Cultured With PancreaCult™ Human Show Pancreatic Marker Expression Levels Similar to Exocrine Tissue

Pancreatic duct organoids grown using the PancreaCult™ Human show marker expression levels similar to those observed in exocrine tissue. Analysis by qPCR showed pancreatic duct organoids were enriched for (A) PDX1 (C) CK19 and (F) LGR5 as compared to total pancreatic tissue, demonstrating enrichment of proliferative duct organoids. Comparable expression of (B) SOX9, (D) cystic fibrosis transmembrane receptor (CFTR), and (E) CA2 was observed in pancreatic duct organoids. Expression levels are normalized to TBP and UBC housekeeping genes (ΔCT) and total pancreas for relative expression levels (ΔΔCT).


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