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Background: In Klinefelter's syndrome patients with azoospermia, microscopic testicular sperm extraction (m-TESE) can be proposed as a therapeutic option. Aim of Study: The aim of this study is to assess the sperm retrieval rate in patients with Klinefelter syndrome in King Faisal Specialist Hospital, Riyadh. Methodology: Retrospective, Chart review of 32 patients with Klinefelter syndrome who underwent m-TESE were reviewed and analyzed. All patients had two sets of semen analysis after 3 − 5 days abstinence of ejaculation with further study of semen by in vitro fertilization (IVF) wash. The hormonal analysis was studied. Ultrasonography of testes was assessed preoperatively. Testicular tubules were sent to the IVF laboratory and were studied under the microscope looking for sperms. Some testicular tissues were sent for the histopathology diagnosis Results: Patients' mean age was 34.9 ± 6.0 years. Mean hormonal levels of E2, FSH, LH, prolactin, and testosterone were 96.0 ± 22.0 pmol/L, 29.8 ± 5.4 IU/L, 19.0 ± 2.9 IU/L, 15.4 ± 3.6 ug/L, and 10.0 ± 1.9 nmol/L, respectively. There were two mosaic Klinefelter syndrome patients (6.25%), whereas 30 patients had a nonmosaic form (93.75%). The overall sperm retrieval rate was 37.5%. All patients had small bilateral testes. Sperm retrieval was successful in three patients with hypospermatogenesis, one patient with maturation arrest, and 8 patients with Sertoli-cell-only pattern. Four patients with complete hyalinization of testicular tissues had complete failure to retrieve sperms. The pregnancy rate after intra-cytoplasmic sperm injection was 50%. Conclusions: The sperm retrieval rate in Klinefelter syndrome patients with m-TESE is in accordance with most of those reported in the literature. Regarding histopathology, hypo-spermatogenesis showed a favorable outcome. The pregnancy rate with intra-cytoplasmic sperm injection was 50%.