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Cell Biology JoVE (Open Access) Citable · DOI

Isolation of Retinal Stem Cells from the Mouse Eye

DOI: 10.3791/2209-v
What you'll learn
  • ✓ Isolate retinal stem cells from mouse ciliary epithelium
  • ✓ Culture stem cells into clonal retinal spheres in vitro
  • ✓ Verify self-renewal and multipotentiality of isolated stem cell populations
Protocol

In this video, we will demonstrate how to isolate retinal stem cells from the ciliary epithelium of the mouse eye and grow them in culture to form clonal retinal spheres. The spheres that are isolated possess the cardinal properties of stem cells: self-renewal and multipotentiality.

Difficulty
advanced
Total time
~4–6 hours per mouse (tissue isolation and initial culture setup)
Model organism
Mouse (strain unspecified)
Biosafety
BSL-1

Steps

1
Introduction to retinal stem cell biology and significance

Overview of ciliary epithelium as a source of retinal stem cells and their defining properties of self-renewal and multipotentiality.

▶ 01:05
2
Isolate retinal stem cells from mouse eye

Perform surgical and tissue dissection techniques to extract ciliary epithelium from mouse eye and enzymatically dissociate cells for culture initiation.

▶ 01:21
3
Assess formation of clonal retinal spheres

Culture isolated cells under conditions promoting sphere formation and evaluate sphere characteristics and cellular composition.

▶ 06:27
4
Summarize retinal stem cell isolation outcomes

Review key findings and relevance of the protocol for retinal stem cell research and regenerative medicine applications.

▶ 07:02
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