Skip to content
← Back to browse
PANCREAS Publication-derived

Organoid Models of Human and Mouse Ductal Pancreatic Cancer

Source Boj et al., 2015 · Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), Utrecht; Cold Spring Harbor Laboratory, USA · 10.1016/j.cell.2014.12.021

👤 Sylvia F. Boj, Chang-Il Hwang, Lindsey A. Baker, Hans Clevers, David A. Tuveson ⏱ 20 days 📋 8 phases 🧫 Human pancreatic ductal cells from resected tissue and endoscopic biopsies; Mouse pancreatic ductal cells from wild-type and transgenic strains (KC, KPC mice)

Abstract

This protocol describes the establishment and propagation of three-dimensional pancreatic ductal organoids from normal and neoplastic murine and human pancreatic tissue. The organoids recapitulate pancreatic intraepithelial neoplasia (PanIN) to pancreatic ductal adenocarcinoma (PDA) progression in vitro and in vivo, enabling molecular profiling and identification of disease-stage-specific biomarkers and therapeutic targets.

Cell source
Human pancreatic ductal cells from resected tissue and endoscopic biopsies; Mouse pancreatic ductal cells from wild-type and transgenic strains (KC, KPC mice)
Application
Disease modeling; Pancreatic cancer progression modeling and therapeutic target discovery

Protocol overview

31 steps across 8 phases

Tissue Harvest and Preparation Day 0
  1. 1 Collect pancreatic tissue
  2. 2 Tissue mincing (for tumors and metastases)
Enzymatic Digestion Day 0–1
  1. 1 Digest mouse normal and preneoplastic ductal tissue
  2. 2 Digest murine tumor and metastatic tissue
  3. 3 Digest human normal pancreatic tissue
  4. 4 Digest human tumor tissue
Organoid Establishment in Matrigel Day 0–1
  1. 1 Prepare Matrigel suspension for murine ducts
  2. 2 Prepare Matrigel suspension for human tissue
  3. 3 Plate in culture wells
Culture Medium Preparation and Application Day 0
  1. 1 Prepare murine organoid culture medium
  2. 2 Prepare human normal (hN) organoid culture medium
  3. 3 Prepare human tumor (hT) organoid culture medium
  4. 4 Add culture medium to wells
Organoid Outgrowth and Passaging Day 1–20+
  1. 1 Observe organoid outgrowth
  2. 2 Change medium every 3–4 days
  3. 3 Passage murine organoids
  4. 4 Passage human normal organoids
  5. 5 Passage human tumor organoids
Cryopreservation Any passage
  1. 1 Prepare organoids for cryopreservation
  2. 2 Freeze organoids gradually
  3. 3 Thaw organoids for use
Genetic Manipulation (Optional - Adenoviral Cre Infection for Kras Activation) Passage 3 onward
  1. 1 Prepare organoids for adenoviral infection
  2. 2 Infect organoids via spinoculation
  3. 3 Culture infected organoids
  4. 4 Assess Cre-mediated recombination
Orthotopic Transplantation (Optional) Passage 1–3+
  1. 1 Prepare organoids for transplantation
  2. 2 Prepare recipient mice for surgery
  3. 3 Perform orthotopic injection into pancreas
  4. 4 Close surgical incision
  5. 5 Post-operative care
  6. 6 Monitor for tumor development

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

This SOP was authored by Organthis based on the published method in Boj et al., 2015. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence