Vitrification and xenografting of human ovarian tissue

Capsule:
Two vitrification protocols were shown to preserve the morphology and viability of human preantral follicles. After warming and xenografting to nude mice for 1 week, these follicles were able to survive and develop.

Authors:
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.

Volume 98, Issue 5, Pages 1291-1298.e2, November 2012

Abstract:

Objective:
To assess the efficiency of two vitrification protocols to cryopreserve human preantral follicles using a xenografting model.

Design:
Pilot study.

Setting:
Gynecology research unit in a university hospital.

Patient(s):
Ovarian biopsies were obtained from 7 women aged 30-41 years.

Intervention(s):
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.

Result(s):
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.

Conclusion(s):
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.

  • Javier Domingo del Pozo

    Congratulations. I think this is very interesting. I would like to ask you a question. Do you have or do you know any experience about the lifespam of the graft using ovarian tissue vitrification and warming instead slow freezing?

    • Thanks, we have been very interested in this topic. As far as I know,
      only a couple of hospitals started grafting vitrified-warmed human
      ovarian tissue. So, for the moment, it is too early to know the graft
      lifespan in the patients. Regarding other animal species, I do not think any of the published studies calculated the duration of the grafted tissue.

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