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. 2017 Dec 6;7:17050. doi: 10.1038/s41598-017-17138-y

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© The Author(s) 2017

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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The effect of PLO on stromal cells. (A) BESC isolated from 3 animals, and (B) HESC from 3 independent passages were challenged for 2 h with control medium (■), or media containing the indicated concentrations of PLO (), and cell viability evaluated by MTT assay. Data are presented as mean (SEM), and analyzed by one-way ANOVA with Dunnett’s multiple comparison post-hoc test; values differ from control, **P < 0.01, ***P < 0.001. (C) BESC isolated from 5 animals, and (D) HESC from 3 independent passages were grown in control serum-free medium or medium containing methyl-β-cyclodextrin for 24 h to deplete cholesterol, and then challenged for 2 h with control medium (■), or media containing PLO () using 100 HU/ml for BESC and 200 HU/ml for HESC, and leakage of LDH was measured in cell supernatants. Data are presented as mean (SEM), and analyzed by one-way ANOVA with Dunnett’s multiple comparison post-hoc test; values differ from control, **P < 0.01. (E) BESC isolated from 3 animals, and (F) HESC from 3 independent passages, were challenged for 24 h with control serum-free medium (■), or media containing PLO () or DS-PLO (DS, ), and cell viability evaluated by MTT assay. Data are presented as mean (SEM) percent viability of control, and analyzed by one-way ANOVA with Dunnett’s multiple comparison post-hoc test; values differ from control, **P < 0.01, ***P < 0.001.

Inline graphic — The effect of PLO on stromal cells. (A) BESC isolated from 3 animals, and (B) HESC from 3 independent passages were challenged for 2 h with control medium (■), or media containing the indicated concentrations of PLO (), and cell viability evaluated by MTT assay. Data are presented as mean (SEM), and analyzed by one-way ANOVA with Dunnett’s multiple comparison post-hoc test; values differ from control, **P < 0.01, ***P < 0.001. (C) BESC isolated from 5 animals, and (D) HESC from 3 independent passages were grown in control serum-free medium or medium containing methyl-β-cyclodextrin for 24 h to deplete cholesterol, and then challenged for 2 h with control medium (■), or media containing PLO () using 100 HU/ml for BESC and 200 HU/ml for HESC, and leakage of LDH was measured in cell supernatants. Data are presented as mean (SEM), and analyzed by one-way ANOVA with Dunnett’s multiple comparison post-hoc test; values differ from control, **P < 0.01. (E) BESC isolated from 3 animals, and (F) HESC from 3 independent passages, were challenged for 24 h with control serum-free medium (■), or media containing PLO () or DS-PLO (DS, ), and cell viability evaluated by MTT assay. Data are presented as mean (SEM) percent viability of control, and analyzed by one-way ANOVA with Dunnett’s multiple comparison post-hoc test; values differ from control, **P < 0.01, ***P < 0.001.