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. 2019 Aug 23;9:12297. doi: 10.1038/s41598-019-48814-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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hiPSC preserve pluripotency throughout four passages. Cells grown as monolayer culture (ML) and suspension culture (SUS) in either mTeSR™1 or mTeSR™3D cell culture medium were investigated for pluripotency status on gene and protein level. (A) The gene expression of pluripotency-associated markers OCT3/4, SOX-2, and NANOG was confirmed by qRT-PCR. (B) Likewise, flow cytometry analysis revealed consistent expression of pluripotency transcription factors. Values in (A) and (B) are normalized to marker expression of pluripotent starting cultures. Error bars indicate mean ± SD, n = 3. (C) PluriTest results (HT12v4 arrays) normalized to hESC H9 cells cultured in E8 and TeSR medium (n = 2 for normalization samples). The background encodes an empirical density map indicating areas of high Pluripotency/low Novelty Scores (red) and high Novelty/low Pluripotency Scores (blue); thresholds for Pluripotency (20, horizontal) and Novelty (1.67, vertical) Scores are indicated with dashed lines. All samples pass both Pluripotency and Novelty Score criteria. (D) Magnification of PluriTest results from (C) hiPSC starter culture is highlighted green.