○孫 こうい1 浅見 真紀2 大田 浩1,2 小阪 美津子1
JST/PRESTO 岡山 インキュべーションセンター1，神戸理研 発生再生研2
Previous our study reported that once differentiated chick IPE cells can be stably maintained in standard control medium and can also transdifferentiate into lens cells when FGF-2 and EGF existed. In this study we investigated whether IPE cells have potency to transdifferentiate into specific neuronal types. Dissociated IPE cells isolated from postnatal chick were cultured under various culture conditions. Cell morphology and the expression of retinal neuron markers were checked by immunostaining and RT-PCR methods. When plated onto laminin-coated dish and cultured in serum-free medium with FGF- 2, the IPE cells lost pigmentation and changed into neuron-like cells. Nearly 50% cells expressed tubulin, and 10% cells expressed GFAP. Some of the cells also expressed the retinal neuronal markers as opsin and protein kinase C, which is the specific marker of photoreceptor and bipolar cells respectively. In addition when the IPE cells were co-cultured with dissociated chick retinal cells from E5 embryos in rotation culture, they joined in the formation of retinal spheroids reconstituting a complete arrangement of all retinal layers. Some of the IPE cells changed into photoreceptor-like cells in the outer layer of the retinal spheroids. Further study has shown that mammalian IPE cells can also transdifferentiate into retinal specific neuron type under appropriate culture conditions. These results indicate that IPE cells from higher vertebrate species have potency to transdifferentiate into retinal specific neurons in vitro.
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