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BRAIN Publication-derived

Meningioma Brain Tumouroids (2024)

Full SOP with Academic or a plan

Source Kim et al., 2024 · The Catholic University of Korea; Seoul St. Mary's Hospital · 10.1186/s12935-024-03225-4

Authors: Dokyeong Kim, Junseong Park, Hyeon-Chun Park, Songzi Zhang, Minyoung Park, Soon A. Park, Sug Hyung Lee, Youn Soo Lee, Jae-Sung Park, Sin-Soo Jeun, Yeun-Jun Chung, Stephen Ahn 7 phases Cell source: Patient-Derived Tissue (Meningioma)

Abstract

This protocol establishes patient-derived meningioma organoids (MNOs) from surgically resected tumor tissue using mechanical mincing without enzymatic dissociation, thereby preserving the tumor microenvironment and diverse stromal cell types. MNOs maintain functionality, morphology, and genetic profiles over long-term culture (>9 weeks) and repeated cryopreservation-recovery cycles, enabling their use as preclinical models for drug screening and precision medicine applications.

Cell source
Patient-Derived Tissue (Meningioma)
Application
Disease modeling and precision medicine; drug screening

Protocol overview

31 steps across 7 phases

Phase 1 · Tissue collection and preparation Day 0
  1. a Tissue collection and initial preservation
  2. b Mechanical tissue mincing
  3. c RBC lysis buffer treatment
Phase 2 · Meningioma organoid establishment and maintenance Days 1-14
  1. a Culture initiation in growth medium
  2. b Medium changes
  3. c Organoid passaging
Phase 3 · Cryopreservation and recovery Variable (after established organoid culture)
  1. a Preparation for cryopreservation
  2. b Transfer to freezing medium and storage
  3. c Thawing and recovery
Phase 4 · Organoid viability and cytotoxicity assay Variable (assay specific; 72 h for drug treatment)
  1. a Single organoid transfer to 96-well plate
  2. b WST viability assay
  3. c LDH cytotoxicity assay setup
  4. d LDH detection
  5. e Positive control treatment
Phase 5 · 3D invasion assay Variable (0-7 days)
  1. a Matrix preparation
  2. b Matrix seeding in wells
  3. c Culture medium overlay and incubation
  4. d Invasion quantification
Phase 6 · Histology and immunostaining Variable
  1. a Organoid fixation and freezing
  2. b Cryostat sectioning
  3. c H&E staining
  4. d Masson's trichrome staining
  5. e Immunohistochemistry (IHC) with DAB peroxidase
  6. f Slide scanning and imaging
  7. g Immunofluorescence (IF) imaging
Phase 7 · Whole-exome sequencing (WES) Variable (offsite analysis typically 2-4 weeks)
  1. a DNA extraction
  2. b Library preparation and exome capture
  3. c Sequencing
  4. d Sequence data analysis and variant calling
  5. e Copy number variant (CNV) analysis
  6. f Variant visualization and comparison

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Attribution

This SOP was written by Organthis based on the published method in Kim et al., 2024. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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