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BRAIN Publication-derived

Novel Human Meningioma Organoids Recapitulating Aggressive Cell Subpopulations

Source Huang et al., 2023 · Sun Yat-Sen University, Department of Neurosurgery, Xiangya Hospital, Central South University · 10.1002/advs.202205525

👤 Meng Huang, Shao Xu, Yuzhe Li, Li Shang, Xiudan Zhan, Chaoyin Qin, Jun Su, Zijin Zhao, Yi He, Lina Qin, Wei Zhao, Wenyong Long, Qing Liu ⏱ 90 days 📋 9 phases 🧫 Patient-Derived Tumor (Meningioma)

Abstract

This protocol describes the establishment of patient-derived meningioma organoid (MO) models that retain intratumoral heterogeneity and the aggressive SULT1E1+ cell subpopulation found in high-grade meningiomas. MOs preserve histological features, genetic mutations, immune cell composition, and biological invasiveness, enabling the identification of SRT1720 as a therapeutic candidate.

Cell source
Patient-Derived Tumor (Meningioma)
Application
Disease modeling and therapeutic target screening for high-grade meningioma

Protocol overview

35 steps across 9 phases

Patient Tumor Sample Preparation and Initial Processing Day 0
  1. 1 Fresh Meningioma Tumor Collection
  2. 2 Tumor Sample Dissection for Organoid Culture
Meningioma Organoid (MO) Culture Setup Day 0–1
  1. 3 Prepare MO Culture Medium Stock
  2. 4 Prepare Complete MO Culture Medium
  3. 5 Establish MO Culture in Ultra-Low Attachment Plates
  4. 6 MO Medium Replacement
MO Characterization (In Vitro) Day 7–30
  1. 7 Monitor MO Growth by 2D Area Measurement
  2. 8 EdU Incorporation Assay for Cell Proliferation
  3. 9 MO Cryopreservation and Biobanking
  4. 10 MO Recovery from Cryopreservation
MO Histology and Immunostaining Day 7–14
  1. 11 MO Fixation and Paraffin Embedding
  2. 12 H&E Staining of MO and Tumor Sections
  3. 13 Immunofluorescence Staining
  4. 14 Immunohistochemistry Quantification
Genomic and Single-Cell Analysis of MOs Day 7–30
  1. 15 Whole-Exome Sequencing (WES) of MOs and Parental Tumors
  2. 16 scRNA-Seq Library Preparation
  3. 17 scRNA-Seq Data Processing and Cell Type Annotation
  4. 18 Copy Number Variation (CNV) Calling and Pseudotime Analysis
  5. 19 Cell-Cell Communication Analysis
Orthotopic Transplantation and In Vivo Assessment Day 30–90
  1. 20 Surgical Preparation and Epidural MO Transplantation
  2. 21 Subdural MO Transplantation
  3. 22 In Vivo MRI Imaging and Assessment
  4. 23 Tissue Harvest and Histological Analysis
  5. 24 Neoangiogenesis Assessment
High-Throughput Drug Screening Day 7–10
  1. 25 Generate Primary Meningioma Cell Lines from MOs
  2. 26 High-Throughput Screening (HTS) of Epigenetic Compounds
  3. 27 Dose-Response Curve Generation
Compound Treatment of Organoids and Efficacy Assessment Day 7–10
  1. 28 SRT1720 Treatment of SULT1E1+ MOs
  2. 29 PS341 and UNC0631 Treatment of MOs
  3. 30 EdU Assay to Quantify Proliferation After Treatment
  4. 31 Immunostaining for SULT1E1 After Drug Treatment
  5. 32 Western Blot Validation of SULT1E1 Downregulation
Combination Therapy: SRT1720 and Radiation Day 8–9
  1. 33 Pre-Treatment with SRT1720
  2. 34 Radiation of SRT1720-Treated MOs
  3. 35 Post-Irradiation EdU Proliferation Assessment

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Huang et al., 2023. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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