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.
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Area of Interest Cancer, Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Organoids, Stem Cell Biology, Transplantation Research
Formulation Category Serum-Free
Data Figures
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.
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.
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.
The CellRaft AIR? system: A novel system enabling organoid imaging, identification, and isolation A. Stern et al. SLAS Discovery 2022 4
Abstract
Three-dimensional (3D) culture systems have been developed that can re-capitulate organ level responses, simulate compound diffusion through complex structures, and assess cellular heterogeneity of tissues, making them attractive models for advanced in vitro research and discovery. Organoids are a unique subtype of 3D cell culture that are grown from stem cells, are self-organizing, and closely replicate in vivo pathophysiology. Organoids have been used to understand tissue development, model diseases, test drug sensitivity and toxicity, and advance regenerative medicine. However, traditional organoid culture methods are inadequate because they are low throughput and ill-suited for single organoid imaging, phenotypic assessment, and isolation from heterogenous organoid populations. To address these bottlenecks, we have adapted our tissue culture consumable and instrumentation to enable automated imaging, identification, and isolation of individual organoids. Organoids grown on the 3D CytoSort? Array can be reliably tracked, imaged, and phenotypically analyzed using brightfield and fluorescent microscopy as they grow over time, then released and transferred fully intact for use in downstream applications. Using mouse hepatic and pancreatic organoids, we have demonstrated the use of this technology for single-organoid imaging, clonal organoid generation, parent organoid subcloning, and single-organoid RNA extraction for downstream gene expression or transcriptomic analysis. The results validate the ability of the CellRaft AIR? System to facilitate efficient, user-friendly, and automated workflows broadly applicable to organoid research by overcoming several pain points: 1) single organoid time-course imaging and phenotypic assessment, 2) establishment of single cell-derived organoids, and 3) isolation and retrieval of single organoids for downstream applications.
A Multiscale Map of the Stem Cell State in Pancreatic Adenocarcinoma. N. K. Lytle et al. Cell 2019
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.
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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. 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 STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
