Purchase this article with an account.
Kenya Yuki, Tetsu Yoshida, Hideyuki Okano, Yoko Ozawa, Kazuo Tsubota; Isolation Of Retinal Progenitor Cells Derived From Mouse iPS Cells Transfected With A Promoter-Rx-DsRed. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2215.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Recent progress in the scientific technology in generating induced pluripotent stem cells (iPS cells) has opened up a new possibility for regenerative medicine. Retinal neural cells, which cannot be replaced by themselves, may be one of the good targets for utilizing the iPS cells. To establish a new therapy, isolation of retinal cells differented from the iPS cells will be an indispensable step. In this study, we discussed the method to purify the retinal progenitor cells derived from mouse iPS (miPS) cells.
A red fluorescent gene, DsRed, was introduced into a gene allele of rax, a marker of retinal progenitor cells, in a BAC clone. The recombinant BAC clone was introduced into the nanog-GFP miPS cells to establish transgenic miPS cell lines. The transgenic miPS cells were cultured under the serum free floating culture of embryoid body-like aggregates combined with Dkk1, Lefty A, serum and activin treatment (SFEB/DLFA) condition for 9 day. The differentiated cells were dissociated into single cells and sorted by fluorescence-activated cell sorting (FACS).
Sixteen lines of transgenic miPS cell lines were obtained. After the SFEB/DLFA-treatment, one transgenic miPS cell line showed 6.36% of Rx-DsRed positive cells, which never overlapped with nanog-GFP.
Rx-positive cells derived from mouse iPS cells were isolated by flow cytometry. This method will help us establishing a better protocol for regenerative therapy, in the future.
This PDF is available to Subscribers Only