Chang Dehui, Li Fudong, Kang Yindong, Yan Yixin, Kong Feiyan, Jiang Wei, Wang Dongxing, Cao Zhigang, Xu Liuting, Huang Chuang, Kang Yafen, Shang Xuejun, Zhang Bin
Department of Urology, The 940th Hospital of the People's Liberation Army Joint Logistics Support Force, Lanzhou, Gansu, China.
Second Department of Surgery, Beijing Fengtai Hospital of Integrated Traditional Chinese and Western Medicine, Beijing, China.
PLoS One. 2025 Feb 10;20(2):e0306235. doi: 10.1371/journal.pone.0306235. eCollection 2025.
To study the effects of L-carnitine and fructose on semen parameters of severe asthenospermia patients by sperm culturing in vitro within 24h. We optimized the energy composition and antioxidant substances of sperm culture medium in vitro (based on Ham's F10 culture medium) by orthogonal test for preparing high quality culture medium. Sperms of 60 patients with idiopathic severe asthenospermia were collected, and cultured in vitro within 24h, by Ham's F10 culture medium added to different concentrations of L-carnitine and fructose and culture temperature, whose effects on sperm motility were observed to determine which is the most appropriate concentration and temperature. For determining the appropriate concentration of L-carnitine and fructose and the suitable culture temperature in Ham's F10 culture medium, the orthogonal experiments were carried out to optimize above three factors, which had great influence on sperm viability, survival rate, deformity rate and DNA fragmentation index (DFI). The final concentration of L-carnitine and fructose was determined in terms of initial tests to assess the effects of different concentrations (4, 8, 12, and 16 mg/ml L-carnitine and 0.125, 0.250, 0.375, and 0.50 mg/ml fructose) on sperm viability and motility in culture. During the operation of processing and culturing sperms in vitro within 24h, orthogonal test showed that sperm viability was better at the final concentration of 8 mg/ml L-carnitine and 0.375 mg/ml fructose in improved Ham's F10 culture medium at 36.5°C. Idiopathic severe asthenospermia sperm can be effectively improved by the modified Ham's F10 culture medium of the final concentration of 8 mg/ml L-carnitine and 0.375 mg/ml fructose at 36.5°C within 24h, which has shown better culture effect and is superior to Ham's F10 basic medium.
通过体外24小时精子培养研究左旋肉碱和果糖对重度弱精子症患者精液参数的影响。我们通过正交试验优化体外精子培养基（基于哈姆氏F10培养基）的能量成分和抗氧化物质，以制备高质量培养基。收集60例特发性重度弱精子症患者的精子，在24小时内进行体外培养，在哈姆氏F10培养基中添加不同浓度的左旋肉碱和果糖以及培养温度，观察其对精子活力的影响，以确定最合适的浓度和温度。为了确定哈姆氏F10培养基中左旋肉碱和果糖的合适浓度以及合适的培养温度，进行了正交试验以优化上述三个对精子活力、存活率、畸形率和DNA碎片指数（DFI）有很大影响的因素。根据初步试验确定左旋肉碱和果糖的最终浓度，以评估不同浓度（4、8、12和16mg/ml左旋肉碱以及0.125、0.250、0.375和0.50mg/ml果糖）对培养中精子活力和运动能力的影响。在24小时内体外处理和培养精子的操作过程中，正交试验表明，在改良的哈姆氏F10培养基中，左旋肉碱最终浓度为8mg/ml、果糖为0.375mg/ml、温度为36.5°C时，精子活力更好。特发性重度弱精子症精子在24小时内，于36.5°C下，采用最终浓度为8mg/ml左旋肉碱和0.375mg/ml果糖的改良哈姆氏F10培养基可得到有效改善，其培养效果更好，优于哈姆氏F10基础培养基。
