COMPARISON OF 4 MECHANICAL METHODS TO RETRIEVE SPERMATOZOA FROM TESTICULAR TISSUE

To maximize the total number of spermatozoa which can be retrieved fro m a testicular biopsy, four mechanical methods for preparation were co mpared, In all, 17 biopsies were divided into four equal fractions, wh ich were weighed individually and prepared by rough shredding (control ), fine mincing, vortexing and crushing (electrical Potter), After res uspension in an erythrocyte-lysing buffer, removal of the remaining ti ssue pieces, washing and centrifugation, the following parameters were evaluated: number of spermatozoa per 100 mg tissue, percentage motili ty, percentage vitality and percentage normal morphology (strict Kruge r criteria), After further purification by discontinuous Percoll centr ifugation, the same parameters were assessed again, This procedure was efficient in obtaining only spermatozoa in the final fraction, Before Percoll centrifugation, only minor differences between methods were o bserved, Percoll centrifugation generally resulted in a pronounced los s of sperm cells, partly because of the abnormal dimensions of the eli minated cells, After Percoll centrifugation, treatment of the testicul ar tissue by fine mincing was the most effective method in terms of th e total number of motile spermatozoa and percentage normal morphology.