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Method Article

Isolating and Culturing Murine Cerebellar Granular Neuron Progenitors

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July 8th, 2025

In This Article

Abstract

Source: Bovio, P., et al., Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark. J. Vis. Exp. (2018)

This video demonstrates the isolation and cultivation of cerebellar neural progenitor cells from a mouse pup's cerebellum. Cerebellar granule neuron progenitors are isolated and purified through differential adhesion, effectively removing astrocytes. These cultured progenitors subsequently differentiate into cerebellar granule neurons.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. Preparations

  1. Preparations for CGNP (cerebellar granular neuron progenitor) isolation
    1. Organize an appropriate mating of mice to generate P5-P7 animals to isolate the cerebellum (3-5 animals per ChIP and condition).
    2. Prepare HBSS/Glucose by adding 6 mg/mL glucose to the HBSS buffer.
    3. Prepare the CGNP cell culture medium (CGM) by adding 1 % (v/v) N2 supplement, 1 % (v/v) Penicillin-Streptomycin-Neomycin, 25 mM KCl and 10 % FCS to ....

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Disclosures

No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
B27 Supplement (50x)Life Technologies17504044Category: Cell culture
Abbreviation/Comment: For CCM
DMEM-F12Life Technologies11320-033Category: Cell culture
Abbreviation/Comment: For CGM
Fetal Bovine Serum 10% (v/v)Gibco10082147Category: Cell culture
Abbreviation/Comment: For CPC isolation and CGM
GlucoseSigma-AldrichG5767Category: Cell culture
Abbreviation/Comment: For CGNP isolation
Glutathione (1.25 mg/ml)Sigma-AldrichG4251Category: Cell culture
Abbreviation/Comment: For CCM
GlycineCarl Roth3187Category: ChIP
Abbreviation/Comment: For cell fixation
Hank's Balanced Salt SolutionLife Technologies14025-100Category: Cell culture
Abbreviation/Comment: HBSS
L-glutamine (200 mM)Life Technologies25030081Category: Cell culture
Abbreviation/Comment: For CCM
LamininSigma-AldrichL2020Category: Cell culture
Abbreviation/Comment: For CPC culturing
N2 supplementLife Technologies17502048Category: Cell culture
Abbreviation/Comment: For CGM
Neurobasal mediumGibco21103049Category: Cell culture
Abbreviation/Comment: For CCM
Penicillin-Streptomycin-Neomycin 1% (v/v)Life Technologies15640055Category: Cell culture
Abbreviation/Comment: PSN, for CCM and CGM
Phosphate buffered salineLife Technologies10010023Category: Cell culture
Abbreviation/Comment: PBS, for CPC isolation
Poly-D-lysineSigma-AldrichP6407Category: Cell culture
Abbreviation/Comment: For CGNP isolation
Poly-L-ornithine hydrobromideSigma AldrichP3655Category: Cell culture
Abbreviation/Comment: For CPC culturing
Potassium chlorideThermo FisherAM9640GCategory: Cell culture
Abbreviation/Comment: KCl, for CGM
Sonic hedgehock (SHH)Sigma-AldrichSRP6004Category: Cell culture
Abbreviation/Comment: For CGNP isolation
Superoxide dismutase (1mg/ml)Sigma-AldrichS7571Category: Cell culture
Abbreviation/Comment: For CCM
Trypsin-EDTA 0,05% (w/v)Sigma Aldrich59417CCategory: Cell culture
Abbreviation/Comment: For CPC isolation

Tags

Cerebellar Granule Neuron ProgenitorsDifferential AdhesionPoly D Lysine CoatingPoly L Ornithine CoatingTrypsin DigestionCell Culture MediaCentrifugationNeubauer Counting ChamberAstrocyte RemovalProgenitor Cell Differentiation