Increased viability of endometrial cells by in vitro treatment with di-(2-ethylhexyl) phthalate  Young-Hoon Kim, M.D., Ph.D., Sung Hoon Kim, M.D., Ph.D., Hoi Woul Lee, M.S., Hee Dong Chae, M.D., Ph.D., Chung-Hoon Kim, M.D., Ph.D., Byung Moon Kang, M.D., Ph.D.  Fertility and Sterility  Volume 94, Issue 6, Pages 2413-2416 (November 2010) DOI: 10.1016/j.fertnstert.2010.04.027 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Viability of endometrial cells in serum-free condition following in vitro DEHP treatment: MTT assay in endometrial cells. Viability of Ishikawa cells according to treatment days (Aa) and after 48 hours (Ab). Viability of endometrial stromal cells according to treatment days (Ac) and after 72 hours (Ad). C: control (vehicle), E: estradiol 10−8 M, P8: DEHP 10−8 M, P6: DEHP 10−6 M. ∗P<.05 versus control, ∗∗P<.01 versus control, ∗∗∗P<.001 versus control. (B) Viability of endometrial cells after exposure to hydrogen peroxide following in vitro DEHP treatment: MTT assay in endometrial cells. Viability of Ishikawa cells according to hydrogen peroxide concetrations (Ba) and after a 4-hour exposure to 300 μM of hydrogen peroxide (Bb). Viability of endometrial stromal cells according to hydrogen peroxide concetrations (Bc) and after a 3-hour exposure to 300 μM of hydrogen peroxide (Bd). C: control (vehicle), E: estradiol 10−8 M, P8: DEHP 10−8 M, P6: DEHP 10−6 M. ∗P<.05 versus control, ∗∗P<.01 versus control, ∗∗∗P<.001 versus control. (C) An FACS analysis after exposure to hydrogen peroxide following in vitro DEHP treatment in Ishikawa cells. Percentage of live cells (Ca), apoptotic cells (Cb), and necrotic cells (Cc) after exposure to 500 μM of hydrogen peroxide. C: control (vehicle), P: DEHP 10−6 M. ∗P<.05 versus control, ∗∗∗P<.001 versus control. (D) An FACS analysis after exposure to hydrogen peroxide following in vitro DEHP treatment in endometrial stromal cells. Percentage of live cells (Da), apoptotic cells (Db), and necrotic cells (Dc) after exposure to 500 and 800 μM of hydrogen peroxide. C: control (vehicle), P: DEHP 10−6 M. ∗P<.05 versus control, ∗∗P<.01 versus control, ∗∗∗P<.001 versus control. (E) Fluorescence microscopic evaluation after Hoechst staining in Ishikawa cells (Ea) and endometrial stromal cels (Eb). C0: control (vehicle) not exposed to hydrogen peroxide, P0: cells treated with DEHP 10−6 M not exposed to hydrogen peroxide, C300/500: control (vehicle) exposed to 300/500 μM of hydrogen peroxide, P300/500: cells treated with DEHP 10−6 M exposed to 300/500 μM of hydrogen peroxide. Fertility and Sterility 2010 94, 2413-2416DOI: (10.1016/j.fertnstert.2010.04.027) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions