首页> 美国卫生研究院文献>Brazilian Journal of Medical and Biological Research >Phytohemagglutinin improves the development andultrastructure of in vitro-cultured goat (Caprahircus) preantral follicles
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Phytohemagglutinin improves the development andultrastructure of in vitro-cultured goat (Caprahircus) preantral follicles

机译:植物血凝素可改善发育和山羊体外培养的超微结构hircus)窦前卵泡

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摘要

The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culturemedium with 10 µg/mL PHA maintains the follicular viability and ultrastructure,and promotes the formation of antral cavity after 6 days of culture invitro.
机译:这项研究的目的是确定植物血凝素(PHA)对体外培养的山羊次要卵泡存活,生长和基因表达的影响。从山羊卵巢皮质中分离出次生卵泡(〜0.2μmm),并在补充有PHA(0、1、10、50、100或200μg)的α-MEM + 中分别培养6天/ mL)。培养6天后,测定卵泡直径和存活率,窦腔形成,超微结构和FSH受体(FSH-R),增殖细胞核抗原(PCNA)和神经元一氧化氮合酶的mRNA表达。所有治疗均维持卵泡存活[α-MEM + (94.59%); 1 µg / mL PHA(96.43%); 10 µg / mL PHA(84.85%); 50 µg / mL PHA(85.29%); 100 µg / mL PHA(88.57%)和200µµg / mL PHA(87.50)],但是与对照组(5.41%)相比,培养基中10µµg / mL PHA的存在增加了胃窦形成率(21.21%)。 ,P <0.05),并确保培养6天后卵母细胞和颗粒细胞超微结构得以维持。与用α-MEM + 培养的卵泡相比,用10μg/ mL PHA培养的卵泡中FSH-R(2.7±0.1)和PCNA(4.4±0.2)的mRNA表达也显着增加。 (1.0±0.1)。总之,文化的补充PHA浓度为10 µg / mL的培养基可维持卵泡活力和超微结构,并在培养6天后促进肛门腔的形成体外。

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