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Method Article

Generating Cortical Pyramidal Neurons from Human Neural Stem Cells

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July 8th, 2025

In This Article

Abstract

Source: Gouder, L., et al,. Three-dimensional quantification of dendritic spines from pyramidal neurons derived from human induced pluripotent stem cells. J. Vis. Exp. (2015)

This video describes a method for preparing glass coverslips coated with polyornithine and laminin to enhance the adhesion and growth of neural stem cells. The technique involves sequential incubations and washes, facilitating the formation of a supportive matrix that promotes neuronal cell growth and differentiation, mimicking structures found in the neurons.

Protocol

  1. Neuronal Culture
    Note: Fibroblast reprogramming in pluripotent stem cells, commitment to the dorsal telencephalon lineage, derivation, amplification, and banking of late cortical progenitors (LCP) were described in Boissart et al. Neuronal differentiation of LCP-like cells was also performed according to Boissart et al with slight modifications. Other procedures have been developed for direct reprogramming of fibroblasts into induced pluripotent stem cells, followed by their differentiation into neurons. This protocol was retained since it allows the selective production of pyramidal glutamatergic neurons.
    1. Treat 6-well culture p....

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Disclosures

No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
PD-PBS (1X), sans Calcium, Magnesium et Phenol RedGibco/ Life Technologies14190169
Poly-L-Ornithine Solution BioreagentSigma AldrichP4957
Mouse lamininDutscher Dominique354232
N2 SupplementGibco/ Life Technologies17502048
B-27 Supplement w/o vit A (50X)Gibco/ Life Technologies12587010
DMEM/NUT.MIX F-12 W/GLUT-IGibco/ Life Technologies31331028
Neurobasal Med SFMGibco/ Life Technologies21103049
2-mercaptoethanolGibco/ Life Technologies31350-010
Pen-SteptomycinGibco/ Life Technologies15140-122
Coverglass 13 mmVWR631-0150
Prolong Gold Antifade Reagent avec DAPIGibco/ Life TechnologiesP36931
Tween(R) 20 Bioextra, Viscous LiquidSigma Aldrich ChimieP7949
Human FibroblastsCoriell Cell Line BiorepositoryGM 4603 and GM 1869Coriell Institute for Medical Research, Camden, NJ, USA
CO2 incubator 
6 well culture plates
Piptette 

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