Presence of spermatogonia in 47,XXY men with no spermatozoa recovered after testicular sperm extraction Dorien Van Saen, M.Sc., Herman Tournaye, M.D.,

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Presence of spermatogonia in 47,XXY men with no spermatozoa recovered after testicular sperm extraction Dorien Van Saen, M.Sc., Herman Tournaye, M.D., Ph.D., Ellen Goossens, Ph.D. Fertility and Sterility Volume 97, Issue 2, Pages 319-323 (February 2012) DOI: 10.1016/j.fertnstert.2011.11.009 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Hematoxylin-eosin staining showing structural integrity of the testicular tissue. (A) Biopsy sample from a 28-year-old patient with KS, showing mainly hyalinized tubules and fibrosis. (B) Biopsy sample from a 43-year-old patient, showing hyalinized tubules and fibrosis. (C) Biopsy sample from a 30-year-old patient with KS, showing degenerated and hyalinized tubules. Degenerated tubules can be recognized by detachment of cells from the basement membrane and a moderately thickened basal lamina. (D) Biopsy sample from a 34-year-old patient; normal tubules are visible (no detachment of cells from the basement membrane and good intercellular contacts). HT = hyalinized tubules; DT = degenerated tubules; NT = normal tubules. ∗Fibrosis. Fertility and Sterility 2012 97, 319-323DOI: (10.1016/j.fertnstert.2011.11.009) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 (A–C) Presence of spermatogonia was evaluated by MAGE-A4 staining. MAGE-A4+ cells were observed in 4 of 22 patients. (A) A 28-year-old patient with KS. Not only spermatogonia were observed but also differentiation up to the spermatocyte level. (B) A 34-year-old patient with KS in whom only spermatogonia were detected. (C) A 24-year-old patient with KS in whom no spermatogonia were observed. (A’–C’) Presence of MAGE-A4+ cells was confirmed by vimentin staining, in which germ cells are visible as negative cells between the positive Sertoli cells. Corresponding vimentin staining was respectively seen in A’–C’. Scale bars represent 100 μm. Fertility and Sterility 2012 97, 319-323DOI: (10.1016/j.fertnstert.2011.11.009) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions