A simple and reliable method for meiotic studies on testicular samples used for intracytoplasmic sperm injection☆

Abstract 

Objective: To develop a reliable and simple method allowing meiotic studies to be performed on testicular samples used for ICSI.

Design: Evaluation of meiotic abnormalities in patients with severe spermatogenic impairment.

Setting: Centre de Médecine de la Reproduction, Marseille.

Patient(s): Two azoospermic men undergoing testicular biopsy for ICSI and one control individual with normal testicular histology.

Intervention(s): The immature germ cells from the patients came from testicular biopsy used for ICSI, after dispersal into a thin cell suspension. Cells were cytocentrifuged to obtain well-spread spermatocytes and then immunocytochemical techniques were performed. We used rabbit polyclonal antibodies against the specific meiotic proteins Cor1 and Syn1 and a human CREST anti-kinetochore antibody.

Main Outcome Measure(s): Synapsis abnormalities in patients with severe spermatogenesis impairment.

Result(s): Pachytene spermatocytes are easily analyzed with this technique, without damage of the axial core and synaptonemal complex. The loss of germ cells is limited.

Conclusion(s): The cytocentrifugation method is the most suitable technique for meiotic studies in patients with severe spermatogenic failure, because it can be used on the testicular cell suspension remaining after ICSI with testicular spermatozoa.

Keywords:  Severe spermatogenic failure, meiosis, cytocentrifugation