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Res Rep Gynaecol Obstet 2017 | Volume 1 Issue 4
November 02-03, 2017 | Chicago, USA
Embryology and In vitro Fertilization
World Congress on
R
estoration of male fertility associated with use of the
cryopreserved testicular tissue would be a significant
advance in human and animal assisted reproductive
technology. The purpose of this study was to test the
effects of four different cryoprotectant agents (CPA) on
spermatogenesis and steroidogenesis in cryopreserved
and allotransplanted neonatal mouse testicular tissue.
Hank’s balanced salt solution (HBSS) with 5% fetal bovine
serum including either 0.7 M dimethyl sulfoxide (DMSO),
0.7 M propylene glycol (PrOH), 0.7 M ethylene glycol (EG),
or glycerol was used as the cryoprotectant solution. Donor
testes were collected and dissected from neonatal pups
of CD-1 mice (one day old). Freezing and seeding of the
testicular whole tissues was performed using an automated
controlled-rate freezer. Four fresh (non-frozen) or frozen-
thawed pieces of testes were subcutaneously grafted onto
the hind flank of each castrated male NCr nude recipient
mouse and harvested after 3 months. Fresh neonatal
testes grafts recovered from transplant sites had the most
advanced rate of spermatogenesis with elongated spermatid
and spermatozoa in 46.6% of seminiferous tubules and had
higher levels of serum testosterone compared to all other
frozen-thawed-graft groups (p<0.05). Fresh grafts and
frozen-thawed grafts in the DMSO group had the highest
rate of tissue survival compared to PrOH, EG, and glycerol
after harvesting (p>0.05). The most effective CPA for the
freezing and thawing of neonatal mouse testes was DMSO
in comparison with EG (p<0.05) in both pre-grafted and
post-grafted tissues based on histopathological evaluation.
Likewise, the highest level of serum testosterone was
obtained from the DMSO CPA group compared to all other
cryoprotectants evaluated (p<0.05). The typical damage
observed in the frozen-thawed grafts included disruption
of the interstitial stroma, intercellular connection ruptures,
and detachment of spermatogonia from the basement
membrane. These findings indicate that neonatal mouse
testes were most effectively preserved when frozen with
HBSS medium with DMSO and that the type of CPA is a
significant factor to obtain the most advanced stages of
spermatogenesis and steroidogenesis after cryopreservation,
thawing, and transplantation of neonatal mouse testes.
e:
cyildiz@mku.edu.trEffect of different cryoprotectant agents on spermatogenesis efficiency in cryopreserved and grafted
neonatal mouse testicular tissue
Yildiz C, Mullen B, Jarvi K, McKerlie C
and
Lo KC
Cengiz Yildiz
University of Mustafa Kemal, Turkey