Skip to content
Back to browse
EPITHELIAL Publication-derived

Head and Neck Squamous Cell Carcinoma Epithelial Organoids

Full SOP with Academic or a plan

Source Driehuis et al., 2019 · Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), University Medical Center Utrecht · 10.1158/2159-8290.cd-18-1522

Authors: Else Driehuis, Sigrid Kolders, Sacha Spelier, Kadi Lõhmussaar, Stefan M. Willems, Lot A. Devriese, Remco de Bree, Emma J. de Ruiter, Jeroen Korving, Harry Begthel, Johan H. van Es, Veerle Geurts, Gui-Wei He, Richard H. van Jaarsveld, Rurika Oka, Mauro J. Muraro, Judith Vivié, Maurice M.J.M. Zandvliet, Antoni P.A. Hendrickx, Nino Iakobachvili, Priya Sridevi, Onno Kranenburg, Ruben van Boxtel, Geert J.P.L. Kops, David A. Tuveson, Peter J. Peters, Alexander van Oudenaarden, Hans Clevers 4 phases Cell source: Human surgical resections and biopsies (normal oral mucosa and HNSCC patient-derived tissue)

Abstract

This protocol describes the establishment and long-term culture of organoids derived from healthy oral mucosa and head and neck squamous cell carcinoma (HNSCC) patient tissue. The organoids recapitulate genetic, molecular, and morphological characteristics of the original tumors and retain tumorigenic potential in vivo, enabling in vitro drug screening for personalized treatment approaches.

Cell source
Human surgical resections and biopsies (normal oral mucosa and HNSCC patient-derived tissue)
Application
Disease modeling; Drug screening for personalized cancer therapy

Protocol overview

17 steps across 4 phases

Phase 1 · Tissue Processing and Digestion Day 0
  1. a Tissue Collection and Preparation
  2. b Enzymatic Tissue Digestion
  3. c Enzyme Inactivation and Filtration
Phase 2 · Organoid Plating and Initial Culture Day 0-1
  1. a BME Suspension Preparation and Plating
  2. b Add Prewarmed Organoid Culture Medium
  3. c Primary Culture Incubation
Phase 3 · Organoid Passaging and Expansion Day 10-14 (first passage); Day 10 onwards (subsequent passages)
  1. a Organoid Harvest and Dissociation
  2. b TrypLE Digestion and Stopping
  3. c Resuspension in Fresh BME and Replating at Split Ratio
  4. d Add Medium and Include ROCK Inhibitor
  5. e Routine Culture Maintenance
Phase 4 · In Vitro Drug Screening Assay Day 0-5 (per drug screen cycle)
  1. a Organoid Preparation for Drug Screening
  2. b Plate Organoids in 384-Well Format
  3. c Add Drug Compounds
  4. d Incubation with Drugs
  5. e Cell Viability Assessment
  6. f Data Analysis and IC50 Calculation

Full SOP

Full SOPs need an account and a plan

Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.

Create an account

Already registered? Log in

Attribution

This SOP was written by Organthis based on the published method in Driehuis et al., 2019. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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