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BRAIN Publication-derived

Patient-Derived Meningioma Organoids: Establishment and Characterization Protocol

Source Zohdy et al., 2025 · Emory University School of Medicine, Department of Neurosurgery · 10.3390/cancers17030526

👤 Youssef M. Zohdy, Arman Jahangiri, Fadi Jacob, Aliaksandr Aksionau, Ali M. Alawieh, Amelia Tong, Bethany Chern, Justin Maldonado, Kimberly Hoang, Edjah Nduom, Brian M. Howard, Daniel L. Barrow, Stewart G. Neill, Zhexing Wen, Gustavo Pradilla, Tomas Garzon-Muvdi ⏱ 45 days 📋 6 phases 🧫 Patient-Derived Tumor (Meningioma)

Abstract

This protocol describes the establishment of three-dimensional meningioma organoids (MEN-Os) directly from patient-resected tumor samples without enzymatic dissociation or exogenous growth factors. MEN-Os preserve tumor architecture, cellular heterogeneity including immune and stromal cells, and recapitulate the transcriptomic and mutational landscapes of original meningiomas, providing a faithful ex-vivo model for studying meningioma biology and evaluating therapeutic strategies.

Cell source
Patient-Derived Tumor (Meningioma)
Application
Disease modeling, Tumor pathophysiology study, Drug screening platform

Protocol overview

37 steps across 6 phases

Patient Tissue Collection and Initial Processing Day 0 (immediate post-resection)
  1. 1 Tissue Collection and Transport
  2. 2 Tissue Quality Assessment
  3. 3 Tissue Dissection and Mincing
  4. 4 Red Blood Cell Lysis
  5. 5 Sample Preparation for RNA Sequencing
  6. 6 Sample Preparation for Histological Analysis
Meningioma Organoid Establishment and Culture Days 1–7
  1. 1 Organoid Medium Preparation
  2. 2 Organoid Plating
  3. 3 Initial Culture Incubation (First 48 Hours)
  4. 4 Medium Change After 48 Hours
  5. 5 Routine Medium Changes (Days 3–28)
  6. 6 Organoid Formation and Growth Monitoring
Organoid Characterization and Analysis Days 7–28
  1. 1 Brightfield Microscopy and Growth Analysis
  2. 2 Sampling for Histological Analysis
  3. 3 Sampling for RNA Sequencing
  4. 4 Hematoxylin and Eosin (H&E) Staining
  5. 5 Immunohistochemistry (IHC) Staining
  6. 6 Immunofluorescence (IF) Staining
RNA Extraction and Sequencing Days 28–35
  1. 1 Sample Preparation and RNA Extraction
  2. 2 RNA Quality Assessment
  3. 3 RNA Storage
  4. 4 RNA-seq Library Preparation
  5. 5 Library Quality Control
  6. 6 RNA Sequencing on Illumina NovaSeq6000
RNA-Sequencing Data Analysis and Bioinformatics Days 36–45
  1. 1 Quality Control of Raw Sequencing Reads
  2. 2 Read Trimming and Adapter Removal
  3. 3 Read Alignment to Reference Genome
  4. 4 Gene Expression Quantification
  5. 5 Differential Gene Expression Analysis
  6. 6 Principal Component Analysis (PCA)
  7. 7 Bulk RNA Deconvolution Analysis
  8. 8 Transcriptomic Mutation Calling
  9. 9 Mutation Similarity Analysis
Statistical Analysis and Quality Control Checkpoints Throughout culture and analysis
  1. 1 Quantitative Data Analysis and Statistical Testing
  2. 2 Documentation of Culture Success and Failure
  3. 3 Verification of Organoid Phenotypic Consistency
  4. 4 Confirmation of Immune Cell Preservation

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Zohdy et al., 2025. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

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