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BRAIN Publication-derived

High-Grade Meningioma Brain Organoids

Full SOP with Academic or a plan

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

Authors: 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 Duration: 90 days 9 phases Cell source: 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

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

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Attribution

This SOP was written by Organthis based on the published method in Huang et al., 2023. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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