Christiani Andrade Amorim, V.M.D., Ph.D., Marie-Madeleine Dolmans, M.D., Ph.D., Anu David, Ph.D., Jonathan Jaeger, M.Sc., Julie Vanacker, B.Sc., Alessandra Camboni, M.D., Ph.D., Jacques Donnez, M.D., Ph.D., Anne Van Langendonckt, Ph.D.
Vol 98, Issue 5, Pages 1291-1298.e2
To assess the efficiency of two vitrification protocols to cryopreserve human preantral follicles using a xenografting model.
Gynecology research unit in a university hospital.
Ovarian biopsies were obtained from 7 women aged 30-41 years.
Ovarian tissue fragments were subjected to one of three cryopreservation protocols (slow-freezing, vitrification protocol 1 and vitrification protocol 2) and xenografted for one week to nude mice.
Main Outcome Measure(s):
The number of morphologically normal follicles after cryopreservation and grafting and fibrotic surface area were determined by histological analysis. Apoptosis was assessed by the TUNEL method. Morphometric analysis of TUNEL-positive surface area was also performed. Follicle proliferation was evaluated by immunohistochemistry.
After xenografting, a difference was observed between the cryopreservation procedures applied. According to TUNEL analysis, both vitrification protocols showed better preservation of preantral follicles than the conventional freezing method. Moreover, histological evaluation showed a significantly higher proportion of primordial follicles in vitrified (protocol 2)-warmed ovarian tissue than in frozen-thawed tissue. The proportion of growing follicles and fibrotic surface area were similar in all groups.
Vitrification procedures appeared to preserve not only the morphology and survival of preantral follicles after one week of xenografting, but also their ability to resume folliculogenesis. In addition, vitrification protocol 2 had a positive impact on the quiescent state of primordial follicles after xenografting.
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