Method Article

Differentiation of Neural Stem Cells to Neurons Using a Compartmentalized Microfluidic Device

July 8th, 2025

In This Article

Abstract

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Source: Paranjape, S. R., et al. Compartmentalization of Human Stem Cell-Derived Neurons within Pre-Assembled Plastic Microfluidic Chips. J. Vis. Exp. (2019)

The video demonstrates the differentiation of neural stem cells (NSCs) using a microfluidic device comprising a somatic and an axonal compartment separated by a microgroove barrier. NSCs are introduced into the somatic compartment and are allowed to adhere. The NSCs differentiate into neurons inside the somatic compartment and extend axons through the microgroove barrier to the axonal compartment.

Protocol

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NOTE: Figure 1A, B shows a schematic of the pre-assembled cyclic olefin copolymer (COC) chip, including the locations of the main channels or compartments, wells, and microgrooves. The compartmentalized chip can establish distinct fluidic microenvironments within a compartment as demonstrated by the isolation of food coloring dye (Figure 1C).

1. Coating of multi-compartment chips for hSC-neurons

NOTE: Figure 2A shows an overview of the coating procedures.

  1. Dissolve poly-L-ornithine in cell culture-gra....

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Results

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Microfluidic device diagram, labeled wells and channels, for cell culture and separation studies.
Figure 1: The pre-assembled COC, multicompartment microfluidic chip. (A) A drawing of the chip identifying the upper and lower wells. The size of the chip is 75 mm x 25 mm, the size of standard microscope slides. (B) A zoo.......

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Disclosures

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No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
complete neural stem cell media:
REC HU EGF
10 UG BIOSOURCE (TM)		ThermoFisher ScientificPHG031420ng/mL
REC HU FGF BASIC
10 UG BIOSOURCE (TM)		ThermoFisher ScientificPHG002420ng/mL
GlutaMAX Supplement (100X)ThermoFisher Scientific350500612mM
KnockOut DMEM/F-12ThermoFisher Scientific12660012
StemPro Neural SupplementThermoFisher ScientificA10508012%
Gibco DPBS without Calcium and MagnesiumThermoFisher Scientific14190144
GIBCO HUMAN NSC (H9) KIT COMBO KITGibcoN7800200
Gibco LamininThermoFisher Scientific23017015
Glass Pasteur pipettesSigma-AldrichCLS7095D5X SIGMA5.75 in length
H9-DERIVED HU NEURAL STEM CELL
1E6 CELLS/VIAL; 1 ML		ThermoFisher Scientific510088
hibernate-E MediumThermoFisher ScientificA1247601
Incubator, 5% CO2 37 °C
Mr. FrostyThermoFisher Scientific5100-0001
Neural differentiation mediaPer 100 mL.
Antibiotic-Antimycotic (100x)ThermoFisher Scientific152401121mL (100X)
Ascorbic acidSigma AldrichA8960200mM
BDNFThermoFisher ScientificPHC707440 ng/mL
Gibco B27 Plus Supplement (50X)FisherScientificA35828012mL (50X)
Gibco CultureOne Supplement (100X)FisherScientificA33202011mL (100X)
Gibco Neurobasal Plus MediumFisherScientificA3582901
StemPro Accutase Cell Dissociation ReagentThermoFisher ScientificA1110501
XC pre-coatXona Microfluidics, LLCXC Pre-Coatincluded with XonaChips
XonaChipXona Microfluidics, LLCXC450450 µm length microgroove barrier
Humidifier TrayXona Microfluidics, LLChumidifier tray

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Tags

Somatic CompartmentAxonal CompartmentMicrogroove BarrierCell DifferentiationNeuron DifferentiationAxon ExtensionPoly l ornithine CoatingLaminin Working Solution

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