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BRAIN Publication-derived

Brain Organoids for Drug Screening and Development

Full SOP with Academic or a plan

Source Kelava et al., 2016 · MRC Laboratory of Molecular Biology, Cambridge · 10.1016/j.stem.2016.05.022

Authors: Iva Kelava, Madeline A. Lancaster Duration: 100 days 5 phases Cell source: Human ESC, Human iPSC

Abstract

This protocol review describes in vitro methods for generating human brain tissue from pluripotent stem cells, including neural rosette approaches, serum-free embryoid body (SFEB/SFEBq) methods, and three-dimensional brain organoids. These systems recapitulate key aspects of human brain development including neural induction, regional patterning, progenitor zone organization, and neurogenesis.

Cell source
Human ESC, Human iPSC
Application
Developmental study, Disease modeling, Drug screening

Protocol overview

15 steps across 5 phases

Phase 1 · Neural Induction via Embryoid Body (EB) Formation 0-7 days
  1. a Generate Embryoid Bodies from Pluripotent Stem Cells
  2. b Culture EBs Under Serum-Free Conditions
Phase 2 · Neural Rosette Formation (2D Method) 7-15 days
  1. a Plate EBs on Coated Dishes
  2. b Maintain and Propagate Rosettes
  3. c Assess Rosette Quality and Cell Composition
Phase 3 · SFEBq Method: Large Rosettes in Semi-3D Culture 7-50 days
  1. a Extend EB Maturation in 3D with Inductive Signals
  2. b Plate Large Aggregates on Coated Dishes
  3. c Continue Differentiation with Growth Factor Support
Phase 4 · 3D Brain Organoid Generation 0-45 days
  1. a Generate and Culture Embryoid Bodies for Organoid Formation
  2. b Embed EBs in Matrigel
  3. c Culture Organoids in 3D Floating Culture with Agitation
  4. d Perform Medium Changes and Monitor Growth
Phase 5 · Quality Control and Characterization Throughout culture (10-100 days)
  1. a Immunofluorescence Staining and Imaging
  2. b RNA Extraction and Gene Expression Analysis
  3. c Assess Tissue Morphology and Regional Identity

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Attribution

This SOP was written by Organthis based on the published method in Kelava et al., 2016. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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