Developmental Competence of Domestic Cat Vitrified Oocytes in 3D Enriched Culture Conditions
Developmental Competence of Domestic Cat Vitrified Oocytes in 3D Enriched Culture Conditions
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DOI：
10.3390/ani9060329
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发表时间：
2019-06-01
期刊：
影响因子：
3
通讯作者：
Luvoni, Gaia Cecilia
中科院分区：
文献类型：
--
作者：
Colombo, Martina;Morselli, Maria Giorgia;Luvoni, Gaia Cecilia
简单总结 卵母细胞玻璃化冷冻是一种冷冻保存方法，可保证人类和野生或家畜遗传物质和生育潜力的长期保存。然而，在家猫中，未成熟玻璃化卵母细胞的体外胚胎发育尚不令人满意。在本研究中，三维（3D）培养系统用于玻璃化卵母细胞的体外胚胎生产，以提供更类似于体内微环境的条件，并且为了进行比较，对照玻璃化卵母细胞在二维（2D）条件下培养。胚胎培养 7 天并评估其发育情况。结果表明，3D富集培养系统能够维持玻璃化卵母细胞的体外成熟和随后的胚胎发育，但与2D系统没有发现差异，仍需要改进以促进玻璃化卵母细胞的发育。 摘要 冷冻损伤严重影响玻璃化卵母细胞（VO）在升温后发育成胚胎的能力。使用提供物理和化学支持并类似于卵母细胞发育的体内微环境的培养条件（例如 3D 支架和共培养系统）可能有助于改善 VO 结果。在本研究中，采用 3D 海藻酸钡微胶囊富集培养系统用于家猫 VO 的体外胚胎生产。冷冻玻璃化加热的卵母细胞在 3D 系统中体外成熟 24 小时，共培养有或没有新鲜卵丘-卵母细胞复合物 (COC)，而对照组的 VO 则在传统的 2D 微滴培养基中培养。体外受精后，根据成熟条件在 3D 或 2D 系统中培养推定胚胎。玻璃化卵母细胞能够在 3D 微胶囊 (17.42 +/- 11.83%) 以及 2D 微滴 (14.96 +/- 8.80%) 中成熟并发育成胚胎，但与 3D 中的伴随 COC 共培养导致 VO 胚胎发育比例相似 (18.39 +/- 16.67%；p = 1.00)，尽管存在 COC允许囊胚形成 (0.95 +/- 2.52%)。总之，直到发育后期的胚胎都是从猫的 VO 中获得的，3D 微胶囊与 2D 微滴相当，但仍需要改善升温后的条件。
Simple Summary Oocyte vitrification is a cryopreservation method that guarantees the long-term conservation of genetic material and fertility potential in humans and wild or domestic animals. However, in the domestic cat the in vitro embryo development of immature vitrified oocytes is not yet satisfactory. In this study, a three-dimensional (3D) culture system was used for the in vitro embryo production of vitrified oocytes to provide conditions more similar to those of the in vivo microenvironment, and for comparison, control vitrified oocytes were cultured in two-dimensional (2D) conditions. Embryos were cultured for seven days and their development was assessed. The results showed that the 3D enriched culture system was able to sustain the in vitro maturation and the subsequent embryo development of vitrified oocytes, but no differences were found with the 2D system and improvements to enhance the development of vitrified oocytes are still needed.Abstract Cryoinjuries severely affect the competence of vitrified oocytes (VOs) to develop into embryos after warming. The use of culture conditions that provide physical and chemical support and resemble the in vivo microenvironment in which oocytes develop, such as 3D scaffolds and coculture systems, might be useful to improve VOs outcomes. In this study, an enriched culture system of 3D barium alginate microcapsules was employed for the in vitro embryo production of domestic cat VOs. Cryotop vitrified-warmed oocytes were in vitro matured for 24 h in the 3D system with or without fresh cumulus-oocyte complexes (COCs) in coculture, whereas a control group of VOs was cultured in traditional 2D microdrops of medium. After in vitro fertilization, presumptive embryos were cultured in 3D or 2D systems according to the maturation conditions. Vitrified oocytes were able to mature and develop into embryos in 3D microcapsules (17.42 +/- 11.83%) as well as in 2D microdrops (14.96 +/- 8.80%), but the coculture with companion COCs in 3D resulted in similar proportions of VOs embryo development (18.39 +/- 16.67%; p = 1.00), although COCs presence allowed for blastocyst formation (0.95 +/- 2.52%). In conclusion, embryos until late developmental stages were obtained from cat VOs, and 3D microcapsules were comparable to 2D microdrops, but improvements in post-warming conditions are still needed.