Cerialle Paula M A, Almodin Carlos G, Radaelli Moacir R M, Minguetti-Câmara Vânia C, Souza Michelle C, Oliveira Carlos A M, Gonçalves Antonio J
Materbaby - Human Reproduction and Genetics, Maringá, Brazil.
Ingá University Center- UNINGA, Maringá, Brazil.
JBRA Assist Reprod. 2017 Jun 1;21(2):73-78. doi: 10.5935/1518-0557.20170019.
This study aimed to compare heterologous to homologous transplantation of fresh ovarian germ cells in rabbits.
Twelve female white New Zealand rabbits (Oryctolagus cuniculus) were randomly numbered and submitted to bilateral oophorectomies. The ovaries from the six odd-numbered rabbits were dissected and cortical germinal tissue was digested in collagenase type 1 to obtain six solutions containing stromal and germ cells, which were injected in the abdominal region of the odd-numbered rabbits themselves (homologous transplantation) and of the even-numbered rabbits (heterologous transplantation) off immunosuppression. Sixty days after transplantation, the tissue around the transplanted region was excised, processed and sent to histological analysis with hematoxylin-eosin staining and Bcl-2 immunohistochemistry to verify the presence and viability of the transplanted cells.
The analyzed specimens contained ovarian stroma, while follicular cells were found in 66.6% of the homologous and in 60% of the heterologous transplant specimens. Mild inflammatory reaction was observed in all heterologous specimens, and in only one (16.7%) of the homologous specimens. However, this inflammatory reaction was not so intense as to cause the death of the implanted cells. Except for the specimens from rabbits 7 and 8, all specimens were stained for Bcl-2, indicating that most of them were viable.
The results of this study supported the viability of heterologous transplantation of fresh ovarian germ cells. However, more studies are required to further our understanding and improve the germ cell separation technique.
本研究旨在比较兔新鲜卵巢生殖细胞的异种移植与同种移植。
将12只雌性白色新西兰兔(穴兔)随机编号并进行双侧卵巢切除术。解剖6只奇数编号兔子的卵巢,将皮质生发组织用1型胶原酶消化,以获得6份含有基质细胞和生殖细胞的溶液,分别注射到奇数编号兔子自身腹部区域(同种移植)和偶数编号兔子腹部区域(异种移植,未进行免疫抑制)。移植60天后,切除移植区域周围的组织,进行处理并送去进行苏木精-伊红染色和Bcl-2免疫组化的组织学分析,以验证移植细胞的存在和活力。
分析的标本中含有卵巢基质,在66.6%的同种移植标本和60%的异种移植标本中发现了卵泡细胞。在所有异种移植标本中均观察到轻度炎症反应,而在同种移植标本中仅1例(16.7%)出现。然而,这种炎症反应并不强烈,不会导致植入细胞死亡。除了来自兔子7和8的标本外,所有标本均进行了Bcl-2染色,表明大多数标本中的细胞是有活力的。
本研究结果支持新鲜卵巢生殖细胞异种移植的可行性。然而,需要更多研究来进一步加深我们的理解并改进生殖细胞分离技术。