PancreaCult™ Organoid Growth Medium (Mouse)
Cell culture medium for establishment and maintenance of mouse pancreatic exocrine organoids
- Dispase (1 U/mL)
1 U/mL dispase in DMEM/F-12
- Collagenase Type IV (1 mg/mL)
Cell dissociation reagent
- Anti-Adherence Rinsing Solution
Rinsing solution for cultureware to prevent cell adhesion
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.
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.
Protocols and Documentation
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PancreaCult™ Organoid Growth Medium (Mouse)
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 this product for growing organoids for drug screening aiming for commercial gain or for other commercial purposes. Purchasers wishing to use the product for purposes other than research use should contact HUB to obtain a further license (an “Organoid-Growth License”). Purchasers may apply for an Organoid-Growth License from HUB and such license 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.
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