Nguyen Thanh Van, Do Lanh Thi Kim, Nguyen Ngoc-Anh Thi, Kikuchi Kazuhiro, Somfai Tamas, Otoi Takeshige
Department of Animal Theriogenology and Surgery, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi, Vietnam.
The Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi, Japan.
Vet Med Int. 2023 Apr 17;2023:5702970. doi: 10.1155/2023/5702970. eCollection 2023.
The aim of this study was to improve the production efficiency of Vietnamese native Ban pig embryos using somatic cell nuclear transfer (SCNT). Fibroblast cells from Ban pigs were injected into the enucleated cytoplasts of crossbred gilts, and the reconstructed embryos were subsequently cultured. In the first experiment, cytoplasts were isolated from oocytes matured in either a defined porcine oocyte medium (POM) or in TCM199 medium supplemented with porcine follicular fluid. Both media were supplemented with gonadotropic hormones, either for the first 22 h of maturation (IVM) or for the entire 44 h of IVM. In the second experiment, the reconstructed SCNT embryos were cultured with or without 50 M chlorogenic acid (CGA). Furthermore, this study examined parthenogenetic embryos. The IVM medium and duration of hormone treatment did not affect embryo development. CGA supplementation to the culture medium significantly increased blastocyst formation rates in parthenogenetic embryos but not in SCNT embryos. However, CGA supplementation significantly reduced the apoptotic index in blastocysts regardless of embryo source. In conclusion, the IVM method did not affect SCNT embryo production, while CGA supplementation during embryo culture improved the quality of SCNT embryos in indigenous pig breeds.
本研究的目的是利用体细胞核移植(SCNT)提高越南本地盘猪胚胎的生产效率。将来自盘猪的成纤维细胞注入杂交后备母猪去核的细胞质中,随后对重构胚胎进行培养。在第一个实验中,从在限定的猪卵母细胞培养基(POM)或添加了猪卵泡液的TCM199培养基中成熟的卵母细胞中分离细胞质。两种培养基都添加了促性腺激素,要么在成熟培养(IVM)的前22小时添加,要么在整个44小时的IVM过程中添加。在第二个实验中,重构的SCNT胚胎在添加或不添加50μM绿原酸(CGA)的条件下进行培养。此外,本研究还检测了孤雌胚胎。IVM培养基和激素处理时间对胚胎发育没有影响。在培养基中添加CGA显著提高了孤雌胚胎的囊胚形成率,但对SCNT胚胎没有影响。然而,无论胚胎来源如何,添加CGA都显著降低了囊胚的凋亡指数。总之,IVM方法不影响SCNT胚胎的生产,而在胚胎培养过程中添加CGA可提高本地猪种SCNT胚胎的质量。