I. Aslam et S. Fishel, SHORT-TERM IN-VITRO CULTURE AND CRYOPRESERVATION OF SPERMATOGENIC CELLS USED FOR HUMAN IN-VITRO CONCEPTION, Human reproduction, 13(3), 1998, pp. 634-638
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).