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Hum. Reprod. Advance Access published online on September 23, 2008
Human Reproduction, doi:10.1093/humrep/den343
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© The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Survival of human pre-antral follicles after cryopreservation of ovarian tissue, follicular isolation and in vitro culture in a calcium alginate matrix

Christiani A. Amorim, Anne Van Langendonckt, Anu David, Marie-Madeleine Dolmans and Jacques Donnez1

Department of Gynecology, Université Catholique de Louvain, Avenue Emmanuel Mounier 52, bte 5247, 1200 Brussels, Belgium

1 Correspondence address. Tel: +32-2-764-95-01; Fax: +32-2-764-95-07; E-mail: jacques.donnez{at}uclouvain.be

BACKGROUND: Ovarian tissue cryopreservation is a promising technique to safeguard fertility in cancer patients. However, in some types of cancer, there is a risk of transmitting malignant cells present in the cryopreserved tissue. To avoid such a risk, pre-antral follicles could be isolated from ovarian tissue and grown in vitro. On the basis of this assumption, the aim of our study was to investigate in vitro survival and growth of pre-antral follicles after cryopreservation of ovarian tissue and follicular isolation, followed by encapsulation in alginate beads.

METHODS: Ovarian biopsies from four patients were frozen and thawed. Pre-antral follicles were then isolated and embedded in an alginate matrix before in vitro culture for 7 days.

RESULTS: Small pre-antral follicles (42.98 ± 9.06 µm) from frozen–thawed tissue can survive and develop after enzymatic isolation and in vitro culture. A total of 159 follicles were incubated in a three-dimensional system (alginate hydrogel), and after 7 days, all of them showed an increase in size (final size 56.73 ± 13.10 µm). The survival rate of the follicles was 90% (oocyte and all granulosa cells viable).

CONCLUSION: Our preliminary results indicate that alginate hydrogels may be a suitable system for in vitro culture of isolated human pre-antral follicles. However, more studies are required to establish whether follicular morphology and functionality can be maintained using this matrix.

Key words: alginate/cryopreservation/follicle/in-vitro culture/ovary

Submitted on March 3, 2008; resubmitted on August 20, 2008; accepted on August 26, 2008.


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