Silva G M, Rossetto R, Chaves R N, Duarte A B G, Araújo V R, Feltrin C, Bernuci M P, Anselmo-Franci J A, Xu M, Woodruff T K, Campello C C, Figueiredo J R
Programa de Pós-Graduação em Ciências Veterinárias (PPGCV),Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais (LAMOFOPA),Universidade Estadual do Ceará (UECE),Av. Paranjana 1700,Campus do Itaperi,CEP: 60740-903,Fortaleza - CE -Brasil.
Laboratory of Manipulation of Oocytes and Preantral Follicles,Faculty of Veterinary Medicine,State University of Ceará,Fortaleza - CE,Brazil.
Zygote. 2015 Aug;23(4):475-84. doi: 10.1017/S0967199414000070. Epub 2014 Mar 26.
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.
本研究的目的是评估二维（2D）和三维（3D）海藻酸盐培养系统对窦前山羊卵泡体外发育的影响。此外，还研究了卵巢供体的生殖年龄对体外培养成功率的影响。从青春期前或成年山羊中分离出窦前卵泡，直接在塑料表面培养（2D）或封装在海藻酸盐基基质中培养（3D）。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系统显示出更高的卵泡存活率、更低的卵母细胞排出率，以及用于IVM和IVF的回收卵母细胞数量更多（P<0.05）。只有成年动物的窦前卵泡产生了MII期卵母细胞和胚胎。在所有测试组中，培养第2天到第12天雌二醇浓度均升高（P<0.05）。相反，3D培养的卵泡中孕酮浓度低于2D培养的卵泡，在培养第2天和第6天存在差异（P<0.05）。我们提供了令人信服的证据，即二维或三维海藻酸盐体外培养系统为实现山羊窦前卵泡在体外完全发育以产生能够受精的成熟卵母细胞和有活力的胚胎提供了一种有前景的方法。
