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BRAIN Publication-derived

Modeling human cortical development in vitro using induced pluripotent stem cells

Source Mariani et al., 2012 · Yale University School of Medicine · 10.1073/pnas.1202944109

👤 Jessica Mariani, Maria Vittoria Simonini, Dean Palejev, Livia Tomasini, Gianfilippo Coppola, Anna M. Szekely, Tamas L. Horvath, Flora M. Vaccarino ⏱ 70 days 📋 4 phases 🧫 Human iPSC

Abstract

This protocol generates three-dimensional multilayered structures from human induced pluripotent stem cells (hiPSCs) that recapitulate early dorsal telencephalic development in vitro. The structures contain polarized radial glia, intermediate neuronal progenitors, and layer-specific cortical neurons with excitatory and inhibitory phenotypes, mimicking the organization of the developing human cerebral cortex at 8–10 weeks postconception.

Cell source
Human iPSC
Application
Developmental study

Protocol overview

9 steps across 4 phases

hiPSC Expansion and Characterization Day 0–prior to differentiation
  1. 1 Culture undifferentiated hiPSC lines
  2. 2 Dissociate hiPSC colonies into single cells
Forebrain Induction and SFEBq Formation Day 0–Day 18
  1. 1 Initiate suspension culture with neural induction factors
  2. 2 Monitor aggregate formation
Aggregate Maturation and Neuronal Differentiation Day 18–Day 45–50
  1. 1 Transfer aggregates to coated surface
  2. 2 Maintain differentiation without additional growth factors
  3. 3 Allow neuronal maturation and layer organization
Synapse Formation and Maturation Day 50–Day 70
  1. 1 Continue culture to day 70 for synaptic maturation
  2. 2 Monitor astroglial differentiation

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Mariani et al., 2012. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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