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精液冷冻贮藏

精液冷冻贮藏(Semen cryopreservation)是指以低温保存精子细胞活性的程序。精液可以透过深低温保存永久保存。若考虑以人类精液进行冷冻贮藏,之后又成功受孕的案例,据文献记载贮藏最长达24年[1]。精液冷冻贮藏可以用在捐精精子银行),可能捐精者和需要精子者相距距离较远,或是在捐精后一段时间才需要精子,若男性接受输精管切除术或是像化学疗法放射线疗法等会影响生育能力的手术前,也可能会先将精液冷冻贮藏,保留日后再养育后代的可能性。在1954年时就已有用冷冻精液解冻后成功受孕的例子[2]

精液冷冻储藏技术广泛应用于男性不孕症患者,及畜牧业的人工繁殖上。对于即将进行化学治疗等细胞毒杀治疗的患者,该技术也能事先保存患者的生殖能力[3]

保存方法

冷冻

最常用的精液冷冻保护剂甘油(占冷冻保护剂溶液的10%),而甘油溶液中也常加入蔗糖、其他的双糖三糖。冷冻保护剂溶液会加入蛋黄或大豆卵磷脂,比较使用这两种物质,精液在解冻后,在精子活动力、变形程度、在体外和玻尿酸盐结合的能力以及其DNA的完整性上,两者没有显著的差异[4]

冷冻保护剂中的其他成分可以增加精子在冷冻后的存活率及受孕率。在冷冻贮藏前,用肝素结合蛋白来处理精子,可以减少精子的冷冻伤害,也减少自由基(Reactive oxygen species)的生成[5]。在冷冻保护剂中加入神经成长因子英语nerve growth factor可以减少精子的死亡速率,增加精子解冻后的活动力[6]。若在冷冻前,用环糊精将胆固醇掺入精子细胞膜中,也可以增加精子活动力[7]

精液的冷冻会用控制速率的慢速冷却(深低温保存,简称SPF),另一种冷却方式是较新式的快速冷却,称为玻璃化(vitrification)。玻璃化的解冻后活动力以及冷冻存活比例都比慢速深低温保存要高[8]

解冻

最理想的解冻温度是40 °C,不过解冻温度的细微调整对精子活动力、顶体状态、ATP含量以及DNA完整性的影响不大[9]。精液冷冻后的解冻也有许多不同的技术,在Di Santo等人2012年的文献中都有提到[10]

再冷冻

比较冷冻再解冻一次和三次的精液,在DNA片段化的风险差异不大。前提是精液和其原始的冷冻保护剂一起再冷冻,中间没有经过精子洗涤或其他类似程序,而用来进行人工受精英语assisted reproduction technology前,会先用密度梯度离心英语density gradient centrifugationswim-up英语swim-up进行分离[11]

对于精子品质的影响

有些证据证实精子在冷冻后,单股断裂、DNA缩合英语DNA condensation片段化的比例增加,这也会增加后代DNA突变的风险。使用抗氧化剂以及良好的冷冻控温可能可以改善此情形[12]

在长期的追踪研究中,用冷冻后精液受孕的人和一般人比较,其先天性障碍染色体畸变的比例相近,还没有证据支持用冷冻后精液受孕的人有较高的风险[12]

历史

对于精液冷冻储藏的历史能够追溯到1776年,意大利人拉扎罗·斯帕兰札尼尝试使用冰雪保存精子[3]。1949年克利斯朵夫·波尔吉利用甘油冷冻保护的特性[13],大幅改善了精液保存技术。自此,人们开始成功以冷冻储藏的精液,进行各种物种的人工受精[3]

相关条目

参考资料

  1. ^ Planer NEWS and Press Releases > Child born after 22 year semen storage using Planer controlled rate freezer 14/10/2004
  2. ^ Fatherhood After Death Has Now Been Proved Possible. Cedar Rapids Gazette. 1954-04-09. 
  3. ^ 3.0 3.1 3.2 Hezavehei, Maryam; Sharafi, Mohsen; Kouchesfahani, Homa Mohseni; Henkel, Ralf; Agarwal, Ashok; Esmaeili, Vahid; Shahverdi, Abdolhossein. Sperm cryopreservation: A review on current molecular cryobiology and advanced approaches. Reproductive BioMedicine Online. 2018-09, 37 (3): 327–339 [2020-02-19]. doi:10.1016/j.rbmo.2018.05.012. (原始内容存档于2022-06-18) (英语). 
  4. ^ Reed ML; et al. Soy lecithin replaces egg yolk for cryopreservation of human sperm without adversely affecting postthaw motility, morphology, sperm DNA integrity, or sperm binding to hyaluronate. Fertility and Sterility. 2009, 92 (5): 1787–1790. doi:10.1016/j.fertnstert.2009.05.026. 
  5. ^ Patel, M., Gandotra, V. K., Cheema, R. S., Bansal, A. K., & Kumar, A. (2016). Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen. Asian-Australasian Journal of Animal Sciences, 29(9), 1247–1255.
  6. ^ Saeednia S, Bahadoran H, Amidi F, Asadi MH, Naji M, Fallahi P, Nejad NA. Nerve growth factor in human semen: Effect of nerve growth factor on the normozoospermic men during cryopreservation process. Iranian Journal of Basic Medical Sciences. 2015, 18 (3): 292–299. 
  7. ^ Purdy PH, Graham JK. Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm. Cryobiology. 2004a, 48: 36–45. doi:10.1016/j.cryobiol.2003.12.001. 
  8. ^ Vutyavanich T, Piromlertamorn W, Nunta S. Rapid freezing versus slow programmable freezing of human spermatozoa. Fertil. Steril. April 2010, 93 (6): 1921–8. PMID 19243759. doi:10.1016/j.fertnstert.2008.04.076. 
  9. ^ Calamera JC, Buffone MG, Doncel GF, et al. Effect of thawing temperature on the motility recovery of cryopreserved human spermatozoa. Fertil. Steril. December 2008, 93 (3): 789–794. PMID 19059590. doi:10.1016/j.fertnstert.2008.10.021. 
  10. ^ Di Santo M, Tarozzi N, Nadalini M, and Borini A. Human Sperm Cryopreservation: Update on Techniques, Effect on DNA Integrity, and Implications for ART (Review). Adv. Urology. 2012, 2012: 854837 [2019-07-12]. PMC 3238352可免费查阅. PMID 22194740. doi:10.1155/2012/854837. (原始内容存档于2018-06-02). 
  11. ^ Thomson LK, Fleming SD, Barone K, Zieschang JA, Clark AM. The effect of repeated freezing and thawing on human sperm DNA fragmentation. Fertil Steril. March 2010, 93 (4): 1147–1156. PMID 19135665. doi:10.1016/j.fertnstert.2008.11.023. 
  12. ^ 12.0 12.1 Kopeika, J.; Thornhill, A.; Khalaf, Y. The effect of cryopreservation on the genome of gametes and embryos: principles of cryobiology and critical appraisal of the evidence. Human Reproduction Update. 2014, 21 (2): 209–227. ISSN 1355-4786. PMID 25519143. doi:10.1093/humupd/dmu063. 
  13. ^ Polge, C.; Smith, A. U.; Parkes, A. S. Revival of Spermatozoa after Vitrification and Dehydration at Low Temperatures. Nature. 1949-10, 164 (4172): 666–666 [2020-02-19]. ISSN 1476-4687. doi:10.1038/164666a0. (原始内容存档于2022-06-18) (英语). 
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精液冷冻贮藏
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