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BRAIN Publication-derived

Brain Organoids for Infection Studies

Full SOP with Academic or a plan

Source Qian et al. · Johns Hopkins University School of Medicine, Baltimore, MD, USA · 10.1016/j.cell.2016.04.032

Authors: Qian X, Nguyen HN, Song MM, Hadiono C, Ogden SC, Hammack C, Yao B, Hamersky GR, Jacob F, Zhong C, Yoon KJ, Jeang W, Lin L, Li Y, Thakor J, Berg DA, Zhang C, Kang E, Chickber M, Nauen D, Ho CY, Wen Z, Christian KM, Shi PY, Maher BJ, Wu H, Jin P, Tang H, Song H, Ming GL Duration: 100 days 4 phases Cell source: Human iPSC

Abstract

Qian et al. developed the SpinΩ miniaturized spinning bioreactor system enabling cost-effective, scalable generation of brain-region-specific organoids including forebrain, midbrain, and hypothalamus tissues from human iPSCs. The platform was applied to model Zika virus infection, demonstrating preferential viral tropism for neural progenitor cells.

Cell source
Human iPSC
Application
Region-Specific Brain Modeling

Protocol overview

12 steps across 4 phases

Phase 1 · iPSC Preparation and Embryoid Body Formation Days 1-7
  1. a iPSC Colony Dissociation
  2. b Transfer Colonies to Ultra-Low Attachment Plate
  3. c Medium Replacement with Induction Medium
Phase 2 · Matrigel Embedding and Pre-patterning Days 7-14
  1. a Embed Organoids in Matrigel
  2. b Mechanically Dissociate from Matrigel
Phase 3 · Spinning Culture and Differentiation Days 14-100
  1. a Transfer to SpinU Bioreactor
  2. b Medium Changes During Growth
  3. c Switch to Maturation Medium at Day 71
  4. d Continue Culture in Maturation Medium
Phase 4 · ZIKV Infection (Optional Disease Model) Day 14, 28, or 80 (24 hours exposure)
  1. a Prepare ZIKV Inoculum
  2. b Apply ZIKV to Organoids
  3. c Replace Medium and Continue Culture

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Attribution

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

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