目的:在我们之前的研究中,我们证实,在卵胞浆内单精子注射过程中,用自体脂肪干细胞(ASC)来源的线粒体补充玻璃化温热的小鼠卵母细胞可增强小鼠受精后发育能力.为了确保这项技术的安全性,我们在小鼠中进行了一项彻底的研究,以调查这种方法形成的后代中潜在的特定畸形的存在。
方法:对补充线粒体后发育的胚胎的建立小鼠进行了跨代比较分析,随后的两代人。繁殖性能,身体生长率,组织病理学参数,血液学参数,日常活动模式,与同龄野生型小鼠相比,评估了这三代雄性和雌性小鼠的每日体温变化。
结果:所有三代中的雄性和雌性动物均显示出与对照组相当的繁殖性能。此外,在所有三个世代中,发现到8周龄的身体生长速率与对照组相当。值得注意的是,在重要器官中未检测到明显的组织病理学异常,包括大脑,心,肝脏,肾脏,肺,卵巢,和睾丸,在研究队列中的任何个体中。血液参数与对照数据一致。在1周的时间内连续监测活动和体温变化(白天和黑夜),揭示了与对照动物非常相似的模式。
结论:将ASC-线粒体注射入卵母细胞可能是一种有前途的技术,可以支持小鼠后代的发育潜能,而不会引起不良的表观遗传事件。然而,在考虑临床应用之前,使用大型动物或非人类灵长类动物进行额外的安全筛查至关重要.
OBJECTIVE: In our previous study, we confirmed that the supplementation of vitrified-warmed murine oocytes with autologous adipose stem cell (ASC)-derived mitochondria during intracytoplasmic sperm injection enhances post-fertilization developmental competence in mice. To ensure the safety of this technology, we conducted a thorough study in mice to investigate the potential presence of specific malformations in offspring developed from this approach.
METHODS: A transgenerational comparative analysis was conducted on founder mice from embryos that developed after mitochondrial supplementation, and two subsequent generations. Reproductive performance, body growth rate, histopathological parameters, hematological parameters, daily activity patterns, and daily body temperature changes in male and female mice across these three generations were assessed in comparison to wild-type mice of the same age.
RESULTS: Both male and female animals in all three generations showed comparable reproductive performance to the control group. Additionally, body growth rate by the age of 8 weeks were found to be comparable to controls across all three generations. Notably, no significant histopathological abnormalities were detected in vital organs, including the brain, heart, liver, kidneys, lungs, ovaries, and testes, in any individuals from the studied cohorts. The blood parameters were consistent with the control data. The continuous monitoring of activity and body temperature changes (both day and night) over a 1-week period revealed a pattern closely resembling that observed in the control animals.
CONCLUSIONS: Injection of ASC-mitochondria into oocytes may be a promising technique to support developmental potential without causing adverse epigenetic events in the offspring in mice. However, before considering clinical application, additional safety screening using larger animals or non-human primates is essential.