Technical Aspects of Laparoscopic Ovarian Autograft in Ewes After Cryopreservation by Slow-Cooling Protocol |
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| Authors: | J Massardier B Courbiere J Lornage C Mazoyer MT Poirel S Martinot M Franck B Salle |
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| Institution: | Hospices Civils de Lyon, Département de Médecine de la Reproduction, Pr. Guérin, Lyon, France;;UniversitéLyon 1, Lyon, France;;UnitéINSERM 418, Dr. Durand, Lyon, France;;Ecole Nationale Vétérinaire de Lyon, Service de Zootechnie, Marcy l'Etoile, France;;Ecole Nationale Vétérinaire de Lyon, Département des Productions Animales, Marcy l'Etoile, France |
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| Abstract: | Iatrogenic ovarian failure and infertility are long term‐term side effects of anticancerous gonadotoxic treatments in children or women of reproductive year. Ovarian cortex cryopreservation can be a solution to preserve immature germinal cells before gonadotoxic treatment, for later transplantation. The aim of our study was to prove the efficiency of a laparoscopic technique for orthotopic graft after a slow‐freezing/thawing protocol, and to evaluate the effect of ovarian cryopreservation and autograft on the primordial follicle survival rate. Experimental surgical study was performed on 6‐ to 12‐month‐old ewes. The study was approved by the ethic committee of the Lyon‐veterinary‐school. The left ovary was removed by laparoscopy and cut in half, and medulla was excised. In group 1 (n = 6), autograft was performed immediately on the right ovary, and in group 2 (n = 6), graft was performed after a slow‐freezing/thawing protocol. The second hemi‐ovary served as an ungrafted control fragment. A polypropylene/polyglactin mesh was included between graft and base to separate the two structures, to help histological analysis. The mean graft performance time was 71 ± 8 min in the first group and 57 ± 10 min in the second. Freezing did not affect the number of primordial follicles. In the ungraft control fragments, the global anomaly rate (cytoplasm plus nuclear anomaly) increased after freezing (p < 0.05). Others results did not reach signification. Pelvic adhesion occurred only once. The post‐graft primordial follicle survival rate was 5.1 ± 2.8% in the non‐frozen group vs. 6.3 ± 2.3% after freezing/thawing. Kruskal–Wallis and Wilkoxon non‐parametric tests were used for statistical analysis. Laparoscopy seems to be a well‐adapted technique for ovarian tissue orthotopic autograft. The main follicle loss occurs before graft revascularization. Our orthotopic graft’s procedure has to be improved to obtain a better graft’s neovascularization, and to have a better long‐term post‐graft primordial follicle survival rate. |
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