Cryopreservation of prepubertal mouse testicular tissue by vitrification.
OBJECTIVE
To compare cryopreservation of prepubertal testicular tissue after vitrification (V) and slow-freezing (SF).


METHODS
Prospective experimental study.


METHODS
Academic research unit.


METHODS
Six-day-old mice.


METHODS
After cryopreservation, viability tests (n = 10) and short-term culture (1 and 3 days) (n = 5) were performed. A comparison was made with fresh (FR) and noncultured controls (FR Ctrl).


METHODS
Tissue viability was assessed by lactate dehydrogenase release assay. Apoptosis (caspase-3) and proliferation (Ki67) were evaluated by immunohistochemistry, and tubular diameter, integrity, and cell density by light microscopy.


RESULTS
Lactate dehydrogenase release was greater after SF than V (54.6% vs. 26.7%), whereas the mean number of apoptotic cells/tubule was higher after V than SF (2.13 vs. 0.07). On day 1, a decrease in cell density was noted in both cryopreserved groups, but this difference was not subsequently observed. On day 3, an increase in proliferation was seen in the SF and V groups versus FR tissue, and similar tubular diameter, integrity, and cell density were found in all cultured groups.


CONCLUSIONS
This study shows that both SF and V protocols preserve survival, development, and integrity of prepubertal mouse testicular tissue in short-term organotypic culture. Additional investigation should now be conducted to assess tissue functionality.
Curaba
Mara
M
Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Verleysen
Magali
M
Amorim
Christiani Andrade
CA
Dolmans
Marie-Madeleine
MM
Van Langendonckt
Anne
A
Hovatta
Outi
O
Wyns
Christine
C
Donnez
Jacques
J
D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D009924:Organ Culture Techniques; D011446:Prospective Studies; D013737:Testis; D014022:Tissue Survival / Q000502:physiology; D058989:Vitrification
D007770:L-Lactate Dehydrogenase