A laboratory modification to testicular sperm preparation technique improves spermatogenic cell yield.

Testicular sperm extraction is a common procedure used to find spermatogenic cells in men with nonobstructive azoospermia. The laboratory processing of biopsied testicular tissues needs to be performed meticulously to acquire a high yield of cells. In this study, the effectiveness of mincing the tissues after testicular biopsy was assessed using histological evaluation, as was the possible adverse effect of residual tissue on the migration of spermatogenic cells during density gradient centrifugation. Our results indicate that testicular residual tissue, when laid on the density gradient medium along with the sperm wash, hinders the spermatogenic cells' forming a pellet during centrifugation, and therefore impairs the intracytoplasmic sperm injection procedure. Whereas the mean number of recovered cells from the sperm wash medium (SWM) with residual tissue is 39.435 ± 24.849, it was notably higher (60.189 ± 28.214 cells) in the SWM without minced tissues. The remaining tissue contained no functional seminiferous tubules or spermatogenic cells in histological sections. In conclusion, the remaining residual tissue after mincing biopsied testicular tissue does not add any functional or cellular contribution to spermatogenic cell retrieval; in fact, it may block the cellular elements in the accompanying cell suspension from migrating through the gradient layers to form a pellet during centrifugation and cause loss of spermatogenic cells.

Unexpected pregnancy despite extremely decreased estradiol levels during ovarian stimulation.

OBJECTIVE: To report two patients with ongoing pregnancies despite a dramatically sharp decrease in E(2) levels after coasting. DESIGN: Case report. SETTING: Reproductive endocrinology and assisted reproduction unit of university hospital. PATIENT(S): One 30-year-old and one 25-year-old woman, both with unexplained infertility, in whom E(2) levels increased up to 6345 and 14,275 pg/mL during ovarian hyperstimulation and decreased by 79.5% and 75.5%, respectively, after coasting. INTERVENTION(S): Two IVF treatments during which coasting was performed after high E(2) levels were observed. IVF cycles were carried out despite abrupt E(2) decrease. MAIN OUTCOME MEASUREMENT(S): Development of ovarian hyperstimulation syndrome (OHSS) or fertilization, cleavage, implantation, and pregnancy rates. RESULT(S): Two embryos (one grade A and one grade B) were transferred into the 30-year-old patient and three embryos (all grade A) were transferred into the 25-year-old patient. Neither woman developed OHSS. Two pregnancies on going at gestational weeks 20 and 14, respectively. CONCLUSION(S): Coasting is practiced to avoid severe complications of ovarian hyperstimulation during IVF cycles and is achieved by withholding gonadotropins. The aim of coasting is to lower E(2) levels to a safer range; however, there has been no consistency with respect to the time of coasting or the safety rates of E(2) decrease. We believe that high rates of E(2) decrease after coasting do not have deleterious effects on implantation.