Home › Cell Biology › One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice
Cell Biology JoVE (Open Access) Citable · DOI

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice

DOI: 10.3791/51225-v
What you'll learn
  • ✓ Isolate neural stem cells from SVZ and DG of individual adult mice
  • ✓ Perform microdissection and tissue dissociation of neurogenic brain regions
  • ✓ Generate both adherent monolayer and neurosphere cultures from single animal
Protocol

Here we describe a detailed protocol for the simultaneous generation of neural precursor cell cultures, as either adherent monolayers or neurospheres, from the subventricular zone and dentate gyrus of individual adult mice.

Difficulty
advanced
Total time
~3–4 hours per mouse (dissection through plating)
Model organism
Mouse (adult, strain unspecified)
Biosafety
BSL-1

Steps

1
Microdissect subventricular zone and dentate gyrus

Extract and isolate SVZ and DG tissue regions from adult mouse brain using fine dissection tools under a stereomicroscope. Tissue is collected in ice-cold dissection medium.

▶ 01:41
2
Dissociate SVZ tissue into single cells

Enzymatically digest SVZ tissue using papain and DNase, then mechanically triturate to generate a single-cell suspension suitable for culture.

▶ 04:13
3
Dissociate dentate gyrus tissue into single cells

Apply the same enzymatic dissociation and trituration protocol to DG tissue to obtain a single-cell suspension.

▶ 05:43
4
Plate cells for monolayer and neurosphere culture

Seed dissociated cells from both regions at defined densities into appropriate culture vessels to initiate either adherent monolayer or floating neurosphere cultures.

▶ 07:36
5
Characterize isolated neural stem cell cultures

Document morphology, growth patterns, and identity of derived monolayer and neurosphere cultures from SVZ and DG over time.

▶ 08:17
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