Max Marilu Constantino, Bizarro-Silva Camila, Búfalo Isabela, González Suellen Miguez, Lindquist Andressa Guidugli, Gomes Roberta Garbelini, Barreiros Thales Ricardo Rigo, Lisboa Lívia Aires, Morotti Fábio, Seneda Marcelo Marcondes
Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Laboratory Animal Reproduction, DCV, CCA, UEL, Londrina, PR, 86051-990, Cx. Postal: 10.011, Brazil.
In Vitro Cell Dev Biol Anim. 2018 Dec;54(10):687-691. doi: 10.1007/s11626-018-0296-9. Epub 2018 Oct 3.
Folliculogenesis is a process of development and maturation of the ovarian follicles, being essential for the maintenance of fertility. In in vivo conditions, 99.9% of the follicles of an ovary do not ovulate and undergo atresia. In order to minimize this loss and to clarify the existing mechanisms, a technique was developed that allows for the in vitro follicular development. The objective of this study was to evaluate the effects of different epidermal growth factor (EGF) concentrations on the in vitro culturing of equine preantral follicles. Ovaries (n = 10) were collected from a local slaughterhouse of mares in seasonal anestrus, washed with 70% alcohol and PBS, and transported. The inner portion of the ovary was divided into 11 fragments of approximately 3 × 3 × 1 mm. A fragment of each ovary was immediately fixed in Bouin (control group). The remaining 10 fragments were individually cultured for 2 and 6 d. The medium was supplemented with different concentrations of EGF (0, 10, 50, 100, and 200 ng/mL). After cultivation, the fragments were processed and classified according to the developmental stage and morphology. In total, 1065 slides containing 6105 tissue sections were evaluated. Within 2 d of culture, there was a higher proportion of intact follicles at the EGF concentrations of 0 and 100 ng/mL (p > 0.05). After 6 d of culture, only the EGF concentration of 100 ng/mL demonstrated a difference when compared to the other treatments (0, 10, 50 and 200 ng/mL of EGF, p > 0.05). There was follicular development after 2 d at all EGF concentrations. Thus, we suggest that EGF promotes follicular survival in equines at a concentration of 100 ng/mL in in vitro cultures of ovarian fragments for 2 d. In addition, we suggest that EGF promotes follicular survival in equines at a concentration of 100 ng/mL in situ cultivation.
卵泡发生是卵巢卵泡发育和成熟的过程，对维持生育能力至关重要。在体内条件下，卵巢中99.9%的卵泡不排卵并发生闭锁。为了尽量减少这种损失并阐明现有机制，开发了一种允许体外卵泡发育的技术。本研究的目的是评估不同表皮生长因子（EGF）浓度对马属动物腔前卵泡体外培养的影响。从当地处于季节性乏情期的母马屠宰场收集卵巢（n = 10），用70%酒精和磷酸盐缓冲液（PBS）冲洗后运输。将卵巢内部切成约3×3×1毫米的11个碎片。每个卵巢的一个碎片立即固定在布安氏固定液中（对照组）。其余10个碎片分别培养2天和6天。培养基中添加不同浓度的EGF（0、10、50、100和200纳克/毫升）。培养后，根据发育阶段和形态对碎片进行处理和分类。总共评估了1065张载玻片，包含6105个组织切片。在培养2天时，EGF浓度为0和100纳克/毫升时完整卵泡的比例较高（p>0.05）。培养6天后，与其他处理（0、10、50和200纳克/毫升的EGF，p>0.05）相比，只有100纳克/毫升的EGF浓度显示出差异。在所有EGF浓度下培养2天后都有卵泡发育。因此，我们建议在卵巢碎片体外培养2天时，EGF以100纳克/毫升的浓度促进马属动物卵泡存活。此外，我们建议在原位培养中，EGF以100纳克/毫升的浓度促进马属动物卵泡存活。
