Risk level of intracytoplasmic spermatid injection for non-mosaic Klinefelter syndrome patients

Abstract

OBJECTIVE: It has been reported that the sex chromosome abnormality incidence rate for ICSI children increased when Klinefelter syndrome patient sperm had been used. We analyzed clinical data of spermatid injections for 99 non-mosaic Klinefelter syndrome patients. DESIGN: Retrospective analysis of clinical data of spermatid injection for 99 non-mosaic Klinefelter syndrome patients. MATERIALS AND METHODS: This study dealt with 99 men who had previously been diagnosed as having non-mosaic Klinefelter syndrome. Many different sites of each testis were biopsied under operation microscope (MD-TESE). Biopsied testicular tissues were in D-PBS containing 0.125% collagenase and 0.01% DNase. Oocytes were transferred to a 295mM solution of mannitol with 0.1mM CaCl2 and 0.05mM Mgcl2 and exposed to an alternating current of 8V/cm 1000KHz for 8s, followed by a single 1200V/cm pulse of direct current for 99μs. Each spermatid, transferred into PVP-HTF, was drawn into an injection pipette (7 μm I.D. for Sa or Sb, 5μm I.D. for Sc, 3μm I.D. for Sd) and the separated nucleus and cytoplasm were injected into the ooplasm. RESULTS: Out of 91 patients, immotile and dead testicular sperms in 12 patients [A: 13.2% (12/91)], differential stages of spermatid in 40 patients [B: 44% (40/91)] and no spermatogenic cells in 4 patients were found using an inverted microscope equipped with Nomalsky optics. Following FISH analysis performed for cases difficult to diagnose, 35 patients showed no existence of haploid cell (spermatid) [38.5% (35/91)] and 3 patients [C: 3.3% (3/91)] had a few them. Pregnancy rates per treatment cycles miscarriage rates and delivery rates in group A, B, C were [A: 27.3% (6/22), 0% (0/6), 27.3% (6/22)], [B: 16.7% (15/90), 40.0% (6/15), 10.0% (9/90)], [C: 33.3% (1/3), 0% (0/1), 33.3% (1/3)] respectively. 16 healthy babies were born (♂:♀= 9:7) with normal karyotype. CONCLUSION: Our data indicates the risk level of intracytoplasmic sperm or spermatid injection into oocytes may be lower than previously expected.