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-obstructive azoospermic men and were cultured in vitro for 96 h as (i) mixed cell populations and (ii) isolated homogeneous populations of primary spermatocytes, round spermatids and elongating spermatids. The cells lost their viability gradually during the first 24 h period. By 72 h almost 90% of the cells were non-viable. Isolated pure fractions showed better viability at each time interval (P < 0.0005). Throughout the culture period primary spermatocytes, elongating spermatids and other non-spermatogenic cells showed no change in their morphology, but almost 22% of round spermatids showed growth of flagella. Most of the round spermatids developed their flagella during the first 4-8 h period of culture. Isolated pure round spermatids showed better flagellar growth compared with mixed cell suspensions (P < 0.0005). The spermatogenic cells were successfully cryopreserved. However, when mixed spermatogenic cell suspensions were cryopreserved, more cells lost their viability compared with when isolated pure fractions were cryopreserved (P < 0.0005).
Aslam
I
I
CARE, The Park Hospital, Arnold, Nottingham, UK.
Fishel
S
S
D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D005307:Fertilization in Vitro; D006801:Humans; D007248:Infertility, Male / Q000628:therapy; D008297:Male; D009845:Oligospermia / Q000473:pathology; D013082:Sperm Tail / Q000502:physiology; D013087:Spermatids / Q000648:ultrastructure; D013094:Spermatozoa / Q000473:pathology / Q000502:physiology*; D013997:Time Factors