Skip to content
← Back to browse
BRAIN Publication-derived

Self-Organization of Polarized Cortical Tissues from Human iPSCs

Source Paşca et al. · Stanford University School of Medicine, Stanford, CA, USA · 10.1038/nmeth.3415

👤 Paşca AM, Sloan SA, Clarke LE, Tian Y, Makinson CD, Huber N, Kim CH, Park JY, O'Rourke NA, Nguyen KD, Smith SJ, Bhatt NP, Barres BA, Paşca SP ⏱ 180 days 📋 5 phases 🧫 Human iPSC

Abstract

Paşca et al. established a Matrigel-free method for generating human cortical spheroids (hCS) from iPSCs using dual SMAD inhibition, producing functional deep- and upper-layer cortical neurons alongside astrocytes over a 90-day culture period. This guided differentiation approach enabled electrophysiological recordings of spontaneous network activity and formed the foundation for the assembloid technology subsequently developed by the Paşca laboratory.

Cell source
Human iPSC
Application
Cortical Development Modeling

Protocol overview

12 steps across 5 phases

hiPSC Culture and Colony Dissociation Day -7 to Day 0
  1. 1 Culture hiPSCs on feeders
  2. 2 Dissociate intact hiPSC colonies
Neural Induction and SMAD Inhibition Day 0 to Day 6
  1. 1 Transfer colonies to low-attachment plates
  2. 2 Apply SMAD signaling inhibitors
Neural Expansion and Progenitor Specification Day 6 to Day 25
  1. 1 Transition to neural medium with growth factors
  2. 2 Perform daily medium changes (days 6-16)
  3. 3 Perform every-other-day medium changes (days 16-25)
Neuronal Differentiation Day 25 to Day 43
  1. 1 Replace growth factors with neurotrophic factors
  2. 2 Maintain BDNF and NT3 supplementation
Maturation and Long-Term Culture Day 43 to Day 180+
  1. 1 Switch to growth-factor-free neural medium
  2. 2 Maintain hCSs through long-term culture
  3. 3 Monitor cellular development milestones

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

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

This wording is awaiting legal review.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence