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. 2016 Sep 2;7(1):127. doi: 10.1186/s13287-016-0380-6

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© The Author(s). 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Sorting for CD59 for RPE purification and stem cell removal. a Use of flow cytometry-based sorting to collect the population expressing CD59 (CD59+) separately from the population not expressing CD59 (CD59–) in a 1:1 mixture of hESCs and RPE cells. b Representative image showing pigmentation in the cell pellets obtained from the CD59+ and CD59– fractions. c Quantitative PCR to measure expression of pluripotency (top) and RPE markers (bottom) in CD59+ and CD59– fractions. The pre-sorted cell suspension is used for comparison. ACT and GAPDH were used as housekeeping genes (n = 4, ± SD). d Representative bright-field images showing the cobblestone architecture of CD59+ and CD59– fractions seeded at a density of 78,000 cells/cm² and cultured for a period of 45 days post sorting. Scale bar = 200 μm. e No CD59-PE fluorescence can be seen in cells described in d as compared with cells freshly stained for CD59 used as a positive control (CD59 pos. ctrl)