Campos-Junior Paulo Henrique Almeida, Alves Thalys Jair Melo, Dias Marco Tulio, Assunçao Carolina Marinho, Munk Michele, Mattos Matheus Silvério, Kraemer Lucas Rocha, Almeida Brígida Gomes, Russo Remo Castro, Barcelos Lucíola, Camargo Luiz Sérgio Almeida, Viana Joao Henrique Moreira
Fertility Preservation Research Group, Department of Natural Sciences, Federal University of Sao Joao Del Rei, Sao Joao Del Rei, Minas Gerais, Brazil, 36301-160.
Laboratory of Genetics and Biotechnology, Department of Biology, Federal University of Juiz de Fora, Juiz de Fora, Minas Gerais, Brazil, 36036-900.
PLoS One. 2016 Jun 30;11(6):e0158109. doi: 10.1371/journal.pone.0158109. eCollection 2016.
Ovarian xenotransplantation is a promising alternative to preserve fertility of oncologic patients. However, several functional aspects of this procedure remained to be addressed. The aim of this study was evaluate the feasibility of xenotransplantation as a strategy to maintain bovine ovarian grafts and produce oocytes. Adult ovarian cortical pieces were xenotransplanted to the dorsal subcutaneous of female NOD-SCID mice (n = 62). Grafts were recovered ten days after xenotransplantation. Host and graft weights; folliculogenesis progression; blood perfusion, relative gene expression and number of macrophage and neutrophil of xenografts; in vitro developmental competence of graft-derived oocytes were evaluated. Folliculogenesis was supported in the grafts, as indicated by the presence of primordial, primary, secondary, antral, and atretic follicles. The xenografts showed a greater volumetric density of atretic follicles and higher hyperemia and number of host-derived macrophage and neutrophil (P<0.05), when compared to non-grafted fragments. There was a higher blood perfusion under the back skin in the transplantation sites of host animals than in control and non-grafted (P<0.01). BAX and PRDX1 genes were up-regulated, while BCL2, FSHR, IGF1R and IGF2R were down-regulated, when compared to the control (P<0.01). Twenty seven oocytes were successfully harvested from grafts, and some of these oocytes were able to give rise to blastocysts after in vitro fertilization. However, cleavage and blastocyst rates of xenograft derived oocytes were lower than in control (P<0.01). Despite showing some functional modifications, the ovarian xenografts were able to support folliculogenesis and produce functional oocytes.
卵巢异种移植是一种有望保留肿瘤患者生育能力的替代方法。然而，该手术的几个功能方面仍有待解决。本研究的目的是评估异种移植作为维持牛卵巢移植物并产生卵母细胞策略的可行性。将成年卵巢皮质片异种移植到雌性NOD-SCID小鼠（n = 62）的背部皮下。异种移植十天后回收移植物。评估宿主和移植物重量、卵泡发生进展、血液灌注、异种移植物的相对基因表达以及巨噬细胞和中性粒细胞数量、移植物来源卵母细胞的体外发育能力。移植物中支持卵泡发生，原始卵泡、初级卵泡、次级卵泡、窦状卵泡和闭锁卵泡的存在表明了这一点。与未移植的片段相比，异种移植物显示出更高的闭锁卵泡体积密度、更高的充血以及更多的宿主来源巨噬细胞和中性粒细胞（P<0.05）。宿主动物移植部位背部皮肤下的血液灌注高于对照和未移植部位（P<0.01）。与对照相比，BAX和PRDX1基因上调，而BCL2、FSHR、IGF1R和IGF2R基因下调（P<0.01）。从移植物中成功收获了27个卵母细胞，其中一些卵母细胞在体外受精后能够发育成囊胚。然而，异种移植来源卵母细胞的卵裂率和囊胚率低于对照（P<0.01）。尽管表现出一些功能改变，但卵巢异种移植物能够支持卵泡发生并产生功能性卵母细胞。
