In vitro development of secondary follicles from pre-pubertal and adult goats cultured in two-dimensional or three-dimensional systems.
In vitro development of secondary follicles from pre-pubertal and adult goats cultured in two-dimensional or three-dimensional systems.
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DOI：
10.1017/s0967199414000070
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发表时间：
2015-08
期刊：
影响因子：
--
通讯作者：
Figueiredo JR
中科院分区：
文献类型：
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作者：
Silva GM;Rossetto R;Chaves RN;Duarte AB;Araújo VR;Feltrin C;Bernuci MP;Anselmo-Franci JA;Xu M;Woodruff TK;Campello CC;Figueiredo JR
本研究的目的是评估二维（2D）和三维（3D）藻酸盐培养系统对山羊腔前卵泡体外发育的影响。此外，还探讨了卵巢供体的生育年龄对体外培养成功率的影响。分离来自青春期前或成年山羊的腔前卵泡，并直接在塑料表面上（2D）或包封在基于藻酸盐的基质中（3D）进行培养。18天后，卵母细胞进行体外成熟（IVM）和体外受精（IVF）以产生胚胎。3D系统在IVM和IVF中显示出较高的卵泡存活率、较低的卵母细胞挤出率和较高的获卵率（P < 0.05）。只有成年动物的腔前卵泡产生MII卵母细胞和胚胎。培养第2 ~ 12天，各组雌二醇浓度均呈上升趋势（P < 0.05）。相反，3D培养的卵泡中孕酮浓度低于2D培养的卵泡，在培养的第2天和第6天差异有统计学意义（P < 0.05）。我们提供了令人信服的证据表明，2D或3D藻酸盐体外培养系统提供了一种有前途的方法，实现山羊腔前卵泡的完全体外发育，以产生能够受精和存活胚胎的成熟卵母细胞。
The aim of this study was to evaluate the influence of two-dimensional (2D) and three-dimensional (3D) alginate culture systems on in vitro development of pre-antral caprine follicles. In addition, the influence of the reproductive age of the ovary donor on the in vitro culture success was investigated. Pre-antral follicles from pre-pubertal or adult goats were isolated and cultured directly on a plastic surface (2D) or encapsulated in an alginate-based matrix (3D). After 18 days, the oocytes underwent in vitro maturation (IVM) and in vitro fertilization (IVF) to produce embryos. The 3D system showed higher rates of follicle survival, lower rates of oocyte extrusion, and a greater number of recovered oocytes for IVM and IVF (P < 0.05). Only pre-antral follicles from adult animals produced MII oocytes and embryos. The estradiol concentrations increased from day 2 to day 12 of culture in all groups tested (P < 0.05). Conversely, progesterone concentrations were lower in 3D-cultured follicles than in 2D-cultured follicles, with differences on days 2 and 6 of culture (P < 0.05). We provide compelling evidence that a 2D or 3D alginate in vitro culture system offers a promising approach to achieving full in vitro development of caprine pre-antral follicles to produce mature oocytes that are capable of fertilization and viable embryos.