Home Cell Biology Targeted Expression of GFP in the Hair Follicle Using Ex Vivo Viral Transduction
Cell Biology JoVE (Open Access) Citable · DOI

Targeted Expression of GFP in the Hair Follicle Using Ex Vivo Viral Transduction

DOI: 10.3791/708-v
What you'll learn
  • Prepare hair follicle histoculture and treat with collagenase for cell dissociation
  • Perform ex vivo viral transduction of hair follicle tissue
  • Visualize GFP expression in transduced follicle stem cells using fluorescence microscopy
Protocol

The hair follicle is a highly complex appendage of the skin containing a multiplicity of cell types. The follicle undergoes constant cycling through the life of the organism including growth and resorption with growth dependent on specific stem cells. The targeting of the follicle by genes and stem cells to change its properties, in particular, the nature of the hair shaft is, discussed. Hair follicle delivery systems are described, such as liposomes and viral vectors for gene therapy. The nature of the hair follicle stem cells is discussed, in particular, its pluripotency.

Difficulty
intermediate
Total time
~4–6 hours (tissue preparation, transduction, and imaging)
Model organism
Human skin tissue (ex vivo hair follicles)
Biosafety
BSL-1

Steps

1
Prepare skin histoculture and apply collagenase treatment

Obtain hair follicle tissue and treat with collagenase to dissociate and isolate follicle structures for downstream transduction. This enzymatic treatment facilitates virus access to target stem cell populations.

▶ 01:16
2
Transduce follicle tissue with viral vector and visualize GFP

Incubate prepared histoculture with viral vector carrying GFP transgene, allowing ex vivo transduction of follicle cells. Image transduced tissue by fluorescence microscopy to confirm GFP expression in target cells.

▶ 03:06
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