NPC Neurosphere Assay: An In Vitro Assay to Assess the Migratory Response of Neural Precursor Cells from the Neurosphere

Published: April 30, 2023

Abstract

Source: Williams, M. et al. Rapid Detection of Neurodevelopmental Phenotypes in Human Neural Precursor Cells (NPCs). J. Vis. Exp. (2018)

This video demonstrates the neural precursor cell neurosphere migration assay—an in vitro assay to study the migratory response of neural precursor cells from the neurosphere—via microscopic analysis.

Protocol

1. NPC Neurosphere Migration Assay Neurosphere Formation Add 1 mL of 100% Expansion Media into a 35 mm dish with no coating substrate. Incubate dishes for at least 15 min at 37 °C before plating NPCs. Prepare 2–3 dishes to ensure that there will be enough neurospheres for the cell migration assay. NOTE: The absence of a coating substrate ensures that NPCs remain suspended in the media, which is essential for neurosphere formatio…

Representative Results

Figure 1: Selecting neurospheres for migration assay. (A) Phase-contrast image of a representative field of neurospheres at 24 h. Spheres are all less than 100 μm and thus, are not collected for the migration assay. (B) Phase-contrast image of a representative field of neurospheres at 72 h. All spheres are within the 100 μm ± 20 µm range, indicating they are ready to be collected…

Açıklamalar

The authors have nothing to disclose.

Materials

Neurobasal Medium  ThermoFischer Scientific 21103049 Component of both NIM and 100% Expansion Medium
PSC Neural Induction Medium: Protocol Link: https://goo.gl/ euub7a  ThermoFischer Scientific   A1647801 This is a kit that consists of Neurobasal (NB) medium and a 50x Neural Induction Supplement (NIS). The NIS is used to make 1X Neural Induction Medium and 100% Expansion Medium
Advanced DMEM/F12 Medium  ThermoFischer Scientific   12634-010 Component of 100% Expansion Medium
hESC-qualified Matrigel Corning  354277 hESC-qualified extracellular matrixmimic gel (ECM-mimic gel)
6-well plates  Corning  COR-3506  Polystyrene plates used for NPC maintenance and for Neurosphere Migration Assay
Human Basic FGF-2  Peprotech 100-18B  Growth factor
Penicillin/Streptomycin  ThermoFischer Scientific   15140122 Antibiotic, component of NIM, 100% Expansion and 30% Expansion Media

Etiketler

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Bu Makaleden Alıntı Yapın
NPC Neurosphere Assay: An In Vitro Assay to Assess the Migratory Response of Neural Precursor Cells from the Neurosphere. J. Vis. Exp. (Pending Publication), e20630, doi: (2023).

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