Table_1_Growth Hormone Promotes in vitro Maturation of Human Oocytes.docx

Increasing the success rate of in vitro maturation (IVM) for human oocytes has a major clinical significance. Previous studies have shown that growth hormone (GH) added into IVM medium could promote IVM of oocytes from non-human beings. However, few studies on systematic IVM for human oocytes with GH have been reported. Human germinal vesicle (GV) oocytes collected for IVM were cultured with different concentrations of GH to optimize the concentration. Metaphase II (MII) stage oocytes obtained from IVM were fertilized by intracytoplasmic sperm injection (ICSI). Maturation rate, fertilization rate, and blastocyst rate were assessed after IVM with or without GH. Furthermore, gene expression profiles were compared in oocytes between the two groups using single-cell RNA-seq. The optimal concentration of GH for IVM was 200 ng/ml, and the maturation rate of this group reached 70% which was double that of the control group (35%, P = 0.004). The fertilization rate (73.1 vs. 60.3%) and blastocyst rate (25.0 vs. 15.5%) both had an increasing trend in the GH group compared to controls. Single-cell RNA-Seq and real-time PCR data showed that GH could significantly enhance the expression of genes associated with meiotic progression and embryo development, such as AURKA (aurora kinase A, P = 0.007), PDIA6 (protein disulfide isomerase family A member 6, P = 0.007), LINGO2 (leucine rich repeat and Ig domain containing 2, P = 0.007), and CENPJ (centromere protein J, P = 0.039). Taken together, GH could promote maturation of human oocytes, probably through accelerating meiotic progression, balancing redox homeostasis of cellular environment, and promoting oocyte developmental competence.

提升体外成熟（IVM）技术在人类卵母细胞中的应用成功率，具有重要的临床意义。既往研究表明，将生长激素（GH）添加至IVM培养基中能够促进非人类生物卵母细胞的体外成熟。然而，关于使用GH进行系统性人类卵母细胞IVM的研究报道尚少。为优化GH浓度，收集用于IVM的人类初级卵母细胞（GV）在一系列不同浓度的GH中进行培养。通过体外成熟获得的减数第二次分裂中期（MII）卵母细胞采用胞浆内精子注射（ICSI）进行受精。评估了IVM后是否添加GH的成熟率、受精率和胚胎囊胚率。此外，利用单细胞RNA测序技术比较了两组卵母细胞之间的基因表达谱。结果显示，GH的最优浓度为200 ng/ml，该浓度组的成熟率达到了70%，是对照组（35%，P = 0.004）的两倍。与对照相比，GH组的受精率（73.1 vs. 60.3%）和胚胎囊胚率（25.0 vs. 15.5%）均呈现上升趋势。单细胞RNA测序和实时PCR数据显示，GH能够显著增强与减数分裂进程和胚胎发育相关的基因表达，例如AURKA（aurora激酶A，P = 0.007）、PDIA6（蛋白二硫键异构酶家族A成员6，P = 0.007）、LINGO2（富含亮氨酸重复序列和Ig结构域蛋白2，P = 0.007）和CENPJ（着丝粒蛋白J，P = 0.039）。综合来看，GH能够促进人类卵母细胞的成熟，可能通过加速减数分裂进程、平衡细胞环境的氧化还原稳态以及提升卵母细胞发育潜能来实现。