April 2009
Volume 50, Issue 13
ARVO Annual Meeting Abstract  |   April 2009
Isolation and Characterization of the Multipotent Neural Crest Stem Cell Derived From Adult Murine Iris Stroma
Author Affiliations & Notes
  • M. Kikuchi
    Ophthalmology, Tohoku University, Sendai, Japan
  • R. Hayashi
    Ophthalmology, Tohoku University, Sendai, Japan
  • S. Kanakubo
    Division of Divelopmental Neuroscience, Tohoku University School of medicine, Sendai, Japan
  • K. Yamamura
    Developmental Genetics, Kumamoto University Institute of Molecular Embryology and Genetics, Kumamoto, Japan
  • N. Osumi
    Division of Developmental Neuroscience, Tohoku University School of Medicine, Sendai, Japan
  • K. Nishida
    Ophthalmology, Tohoku University, Sendai, Japan
  • Footnotes
    Commercial Relationships  M. Kikuchi, None; R. Hayashi, None; S. Kanakubo, None; K. Yamamura, None; N. Osumi, None; K. Nishida, None.
  • Footnotes
    Support  Grants-in-Aid for Scientific Research from ministry of health, labour and welfare, Japan
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1768. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      M. Kikuchi, R. Hayashi, S. Kanakubo, K. Yamamura, N. Osumi, K. Nishida; Isolation and Characterization of the Multipotent Neural Crest Stem Cell Derived From Adult Murine Iris Stroma. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1768.

      Download citation file:

      © ARVO (1962-2015); The Authors (2016-present)

  • Supplements

Purpose: : In the present study, we performed isolation and characterization of multipotent neural crest stem cells from adult murine iris.

Methods: : Iris tissues were isolated from P0-Cre/Floxed-EGFP double transgenic mouse whose neural crest derived tissues were labeled with EGFP. Iris cells were isolated from the iris tissues with enzymatic treatment, and then sphere culture was performed on non-adhesive 96 well-culture dish in serum free medium. Expression of neural crest and stem cell markers in iris spheres were examined by immunofluorescent staining and RT-PCR. To investigate the multipotentiality of these spheres, differentiation assays were performed.

Results: : Microscopic observation showed that EGFP positive cells were localized in iris stroma, trabecular meshwork and corneal endothelium which were generally though to be originated from neural crest. The result of sphere culture showed that almost all the spheres were EGFP positive even though EGFP negative cells existed before sphere culture, indicating neural crest derived-iris stromal cells had high sphere-forming efficiency. Additionally, some of them also had potential to form secondary spheres. Immunofluorescent staining and RT-PCR revealed that the EGFP positive iris spheres expressed neural stem cell markers such as nestin, sox2 and musashi1 as well as neural crest markers such as p75 and AP2. Differentiation assay showed that the sphere forming iris stromal cells had multipotentiality to differentiate into various cell types including neuron, adipocyte and chondrocyte.

Conclusions: : Multipotent stem cells which had neural crest stem cell features were successfully isolated from adult murine iris stroma.

Keywords: iris • cornea: basic science • development 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.