Home › Cell Biology › Isolating Nasal Olfactory Stem Cells from Rodents or Humans
Cell Biology JoVE (Open Access) Citable · DOI

Isolating Nasal Olfactory Stem Cells from Rodents or Humans

DOI: 10.3791/2762-v
What you'll learn
  • ✓ Perform olfactory mucosa biopsy from rodent and human nasal cavities
  • ✓ Isolate and culture multipotent olfactory stem cells from tissue
  • ✓ Differentiate olfactory stem cells into neuronal lineages
  • ✓ Characterize olfactory stem cell properties and sphere formation
Protocol

We describe here a method for biopsying olfactory mucosa from rat and human nasal cavities. These biopsies can be used for either identifying molecular anomalies in brain diseases or isolating multipotent adult stem cells that can be utilized for cell transplantation in animal models of brain trauma/disease.

Difficulty
advanced
Total time
~2–4 hours per biopsy (collection + initial isolation); 7–14 days for sphere formation and differentiation
Model organism
Rat (unspecified strain); Human donors
Biosafety
BSL-1

Steps

1
Collect olfactory mucosa from rat nasal cavity

Perform surgical biopsy of rat olfactory epithelium from the nasal septum under anesthesia. Extract tissue sample containing olfactory mucosa for downstream isolation.

▶ 02:08
2
Collect olfactory mucosa from human nasal cavity

Obtain human olfactory mucosa biopsy from nasal cavity via endoscopic or direct nasal approach. Process tissue under sterile conditions for stem cell isolation.

▶ 03:26
3
Isolate olfactory stem cells from tissue mucosa

Enzymatically digest collected mucosa and mechanically dissociate tissue to release olfactory stem cells. Culture isolated cells in appropriate growth medium.

▶ 04:48
4
Induce sphere formation and neuronal differentiation

Culture isolated olfactory stem cells in conditions promoting neurosphere formation and apply differentiation protocols to generate neuronal progeny.

▶ 06:47
5
Characterize olfactory stem cell phenotype

Analyze marker expression and functional properties of isolated olfactory stem cells to verify multipotency and differentiation capacity.

▶ 07:33
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