Survival of human pre-antral follicles after cryopreservation of ovarian tissue, follicular isolation and in vitro culture in a calcium alginate matrix.
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 microm) 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 microm). The survival rate of the follicles was 90% (oocyte and all granulosa cells viable).


CONCLUSIONS
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.
Amorim
Christiani A
CA
Department of Gynecology, Université Catholique de Louvain, Avenue Emmanuel Mounier 52, bte 5247, 1200 Brussels, Belgium.
Van Langendonckt
Anne
A
David
Anu
A
Dolmans
Marie-Madeleine
MM
Donnez
Jacques
J
D000464:Alginates; D015925:Cryopreservation; D005260:Female; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D009926:Organ Preservation; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology*; D010053:Ovary; D046509:Tissue Culture Techniques
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid