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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.
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© ARVO (1962-2015); The Authors (2016-present)
In the present study, we performed isolation and characterization of multipotent neural crest stem cells from adult murine iris.
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.
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.
Multipotent stem cells which had neural crest stem cell features were successfully isolated from adult murine iris stroma.
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