Praxedes Érica C G, Lima Gabriela L, Bezerra Luana G P, Santos Fernanda A, Bezerra Marcelo B, Guerreiro Denise D, Rodrigues Ana P R, Domingues Sheyla F S, Silva Alexandre R
Laboratory of Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido, BR 110, Km 47, Costa e Silva, 59625-900, Mossoró, RN, Brazil.
Department of Animal Sciences, Instituto Federal de Educação, Ciência e Tecnologia do Ceará (IFCE), CE-292, KM 15, Gisélia Pinheiro, 63115-500, Crato-CE, Brazil.
Reprod Fertil Dev. 2018 Mar;30(3):459-468. doi: 10.1071/RD17051.
The aim of the present study was to evaluate the development of fresh and vitrified agouti ovarian tissue after xenografting to C57Bl/6 severe combined immunodeficiency (SCID) female mice. Ovaries were obtained from five female agoutis and divided into 16 fragments. Five fragments were transplanted immediately to ovariectomised SCID mice and the others were vitrified, stored for 2 weeks and transplanted only after rewarming. Tissue fragments were transplanted under the kidney capsule in recipients. The return of ovarian activity in recipients was monitored by the observation of external signs of oestrus and vaginal cytology over a period of 40 days after transplantation, after which the grafts were removed and evaluated for morphology, cell proliferation and the occurrence of DNA fragmentation. Ovarian activity returned in four of five mice that received fresh ovarian tissue from agoutis and in one of six mice that had received vitrified tissue a mean (±s.e.m.) 20.6±8.6 days after xenotransplantation. After graft removal, a predominance of primordial and primary follicles was observed in all grafts. Vitrification reduced cell proliferation and increased the occurrence of DNA fragmentation in grafted agouti ovarian tissue. In conclusion, the present study demonstrates that xenografted agouti ovarian tissue, fresh or vitrified, is able to promote the return of ovarian activity in ovariectomised SCID C57B1/6 mice. However, improvements to vitrification protocols for agouti ovarian tissue are necessary.
本研究的目的是评估新鲜和玻璃化的刺豚鼠卵巢组织异种移植到C57Bl/6严重联合免疫缺陷(SCID)雌性小鼠后的发育情况。从五只雌性刺豚鼠获取卵巢并分成16个片段。五个片段立即移植到卵巢切除的SCID小鼠中,其他片段进行玻璃化处理,保存2周,仅在复温后移植。组织片段移植到受体的肾包膜下。通过观察移植后40天内发情的外部体征和阴道细胞学来监测受体卵巢活性的恢复,之后取出移植物并评估其形态、细胞增殖和DNA片段化的发生情况。接受新鲜刺豚鼠卵巢组织的五只小鼠中有四只以及接受玻璃化组织的六只小鼠中有一只在异种移植后平均(±标准误)20.6±8.6天恢复了卵巢活性。移植物取出后,在所有移植物中均观察到原始卵泡和初级卵泡占优势。玻璃化处理降低了移植的刺豚鼠卵巢组织中的细胞增殖并增加了DNA片段化的发生。总之,本研究表明,新鲜或玻璃化的异种移植刺豚鼠卵巢组织能够促进卵巢切除的SCID C57B1/6小鼠恢复卵巢活性。然而,有必要改进刺豚鼠卵巢组织的玻璃化方案。