Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, PR China.
Qingdao Longmingniu Embryo Biotechnologies Co. LTD, Qingdao, Shandong 266400, PR China.
Anim Reprod Sci. 2024 Aug;267:107533. doi: 10.1016/j.anireprosci.2024.107533. Epub 2024 Jun 13.
Japanese Black (Wagyu) cattle donors were primed with different protocols and sources of follicle-stimulating hormone (FSH) for successive ovum pickup (OPU) and embryo development after in vitro fertilization (IVF). Following OPU, retrieved cumulus oocyte complexes (COCs) were subjected to IVF, and resulting blastocysts were transferred into recipients to evaluate implantation capability. Experiment 1: The best blastocyst development (45.3 %) and embryo yields (5.0/donor/OPU) were found with oocytes retrieved from donors treated with FSH (STIMUFOL®, Belgium) at a dosage of 150 IU per donor, compared to two others commercial FSH sources. Experiment 2: There were no differences in embryo development or yield with STIMUFOL FSH (total FSH 150 IU/donor) at a priming duration of either 60-h (Regime 1, six FSH injections) or 36-h (Regime 2, four FSH injections). Experiment 3: Compacted COCs required 22-26-h maturation in vitro (IVM) before IVF for optimal blastocyst development (36.1-41.1 %); however, short (18-h) and prolonged (30-h) IVM duration resulted in lower embryonic development. In contrast, expanded COCs resulted in inferior blastocyst development compared to compacted COCs. Immunofluorescence microscopy revealed that the ratio of 89.8 % cumulus compacted COCs were at the germinal vesicle (pachytene) phase while 98.9 % cumulus expanded COCs went through spontaneous meiosis from meiotic metaphase I, anaphase I, telophase I to metaphase II upon OPU retrieval (P<0.05). Pregnancy rates were not different among three FSH sources or different FSH treatments as long as embryos reached the blastocyst stage. Our study found that different sources of FSH used for Wagyu donor priming prior to OPU resulted in differential embryo development potentials, but those embryos that reached out to blastocysts had a competent implantation ability.
日本黑（和牛）牛供体接受了不同的促卵泡激素（FSH）方案和来源的预处理，以便在体外受精（IVF）后进行连续的卵母细胞采集（OPU）和胚胎发育。OPU 后，回收的卵丘-卵母细胞复合物（COCs）进行 IVF，所得囊胚移植到受体中以评估植入能力。实验 1：与其他两种商业 FSH 来源相比，用 FSH（比利时 STIMUFOL®）处理的供体中获得的卵母细胞具有最佳的囊胚发育（45.3%）和胚胎产量（5.0/供体/OPU），剂量为每个供体 150IU。实验 2：用 STIMUFOL FSH（总 FSH 150IU/供体）进行预处理时，无论持续时间为 60 小时（方案 1，六次 FSH 注射）还是 36 小时（方案 2，四次 FSH 注射），胚胎发育或产量均无差异。实验 3：体外受精（IVF）前，致密 COCs 需要 22-26 小时的体外成熟（IVM）才能获得最佳囊胚发育（36.1-41.1%）；然而，短（18 小时）和长（30 小时）的 IVM 时间会导致胚胎发育较低。相比之下，与致密 COCs 相比，扩展 COCs 的囊胚发育较差。免疫荧光显微镜显示，89.8%的卵丘致密 COCs 的卵母细胞处于生发泡（粗线期）阶段，而 98.9%的卵丘扩展 COCs 在 OPU 回收时经历了从减数分裂中期 I、后期 I、末期 I 到中期 II 的自发减数分裂（P<0.05）。只要胚胎达到囊胚阶段，三种 FSH 来源或不同 FSH 处理之间的妊娠率没有差异。我们的研究发现，在 OPU 前对和牛供体进行预处理时使用不同的 FSH 来源会导致胚胎发育潜力的差异，但达到囊胚阶段的胚胎具有良好的植入能力。
