SHORT-TERM IN-VITRO CULTURE AND CRYOPRESERVATION OF SPERMATOGENIC CELLS USED FOR HUMAN IN-VITRO CONCEPTION

Testicular cell suspensions were prepared from obstructive and non-obs tructive azoospermic men and were cultured in vitro for 96 h as (i) mi xed cell populations and (ii) isolated homogeneous populations of prim ary spermatocytes, round spermatids and elongating spermatids. The cel ls lost their viability gradually during the first 24 h period. By 72 h almost 90% of the cells were non-viable, Isolated pure fractions sho wed better viability at each time interval (P < 0.0005). Throughout th e culture period primary spermatocytes, elongating spermatids and othe r non-spermatogenic cells showed no change in their morphology, but al most 22% of round spermatids showed growth of flagella. Most of the ro und spermatids developed their flagella during the first 4-8 h period of culture. Isolated pure round spermatids showed better flagellar gro wth compared with mixed cell suspensions (P < 0.0005), The spermatogen ic cells were successfully cryopreserved, However, when mixed spermato genic cell suspensions were cryopreserved, more cells lost their viabi lity compared,vith when isolated pure fractions were cryopreserved (P < 0.0005).