Sphingosine-1-phosphate suppresses cyclophosphamide induced follicle apoptosis in human fetal ovarian xenografts in nude mice☆ Yuanyuan Meng, M.D., Zhihui.

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Sphingosine-1-phosphate suppresses cyclophosphamide induced follicle apoptosis in human fetal ovarian xenografts in nude mice☆ Yuanyuan Meng, M.D., Zhihui Xu, M.D., Fangfang Wu, M.D., Wenming Chen, M.D., Shuangshuang Xie, M.D., Jun Liu, Ph.D., Xuefeng Huang, M.D., Ying Zhou, M.D. Fertility and Sterility Volume 102, Issue 3, Pages 871-877.e3 (September 2014) DOI: 10.1016/j.fertnstert.2014.05.040 Copyright © 2014 The Authors Terms and Conditions

Figure 1 Histological sections of grafted human fetal ovarian tissue with H&E staining. (A) Ovarian graft exposed to CTX and treated with 2.0 mM S1P (experimental group A). (B) Ovarian graft exposed to CTX and treated with 0.5 mM S1P (experimental group B). (C) Ovarian graft exposed to CTX and treated with PBS (experimental group D). (D) Ovarian graft without CTX exposure (experimental group E). Arrows indicate normal primordial follicles, arrowheads indicate the abnormal follicles. Bar = 100 μm. Fertility and Sterility 2014 102, 871-877.e3DOI: (10.1016/j.fertnstert.2014.05.040) Copyright © 2014 The Authors Terms and Conditions

Figure 2 Representative sections of human fetal ovarian grafts processed for apoptosis using TUNEL staining. (A) CTX + 0.5 mM S1P treated ovarian graft. (B) CTX + 2.0 mM S1P treated ovarian graft. (C) CTX + PET treated ovarian graft. (D) CTX + PBS treated ovarian graft. The greening staining indicates TUNEL positive staining for DNA fragmentation. Bar = 150 μm. Fertility and Sterility 2014 102, 871-877.e3DOI: (10.1016/j.fertnstert.2014.05.040) Copyright © 2014 The Authors Terms and Conditions

Figure 3 Transmission electron microscopy. (A, B) Frozen-thawed ovarian tissue. Arrow indicates a vacuole in the cytoplasm and arrowhead indicates a normal mitochondrion. (C) CTX + 0.5 mM S1P treated ovarian graft. Arrows indicate swollen mitochondria in cytoplasm. (D) CTX + PBS treated ovarian graft. Arrows indicate the abnormal mitochondria with disappeared cristae. (E) Ovarian graft without CTX exposure. Arrows indicate normal mitochondria. (F) CTX + 2.0 mM S1P treated ovarian graft. Arrows indicate normal mitochondria. Fertility and Sterility 2014 102, 871-877.e3DOI: (10.1016/j.fertnstert.2014.05.040) Copyright © 2014 The Authors Terms and Conditions

Supplemental Figure 1 Histologic sections of human fetal ovarian tissue with H&E staining. (A) Fresh human fetal ovarian tissue. (B) Frozen-thawed human fetal ovarian tissue. Arrows indicate primordial follicles. Bar = 100 μm. Fertility and Sterility 2014 102, 871-877.e3DOI: (10.1016/j.fertnstert.2014.05.040) Copyright © 2014 The Authors Terms and Conditions

Supplemental Figure 2 Transplantation of human fetal ovarian tissue. (A) Xenografting of human fetal ovarian tissue beneath the back skin of the nude mice (arrow). (B) The ovarian grafts (arrows) were revascularized and recovered 2 weeks after transplantation. Bar = 5 mm. Fertility and Sterility 2014 102, 871-877.e3DOI: (10.1016/j.fertnstert.2014.05.040) Copyright © 2014 The Authors Terms and Conditions