Tiwari Akansha, Tekcan Merih, Sati Leyla, Murk William, Stronk Jill, Huszar Gabor
Sperm Physiology Laboratory, Yale University, 330 Cedar St, New Haven, CT, 06519, USA.
Medisu Hospital IVF Laboratory, Antalya, Turkey.
J Assist Reprod Genet. 2017 May;34(5):647-657. doi: 10.1007/s10815-017-0888-4. Epub 2017 Mar 9.
Our aim was the development of a safe sperm cryopreservation New Media (NM), composed of consistent and reproducible components devoid of any animal origin, and evaluation of NM in terms of its effect on sperm structure and function as compared to regularly used yolk media (TYM) (Irvine Scientific).
We evaluated patient semen samples and cryopreserved them in duplicates in either NM or TYM. The samples were cryopreserved for either a short term of 1 week or long term of 1 month prior to thawing. The parameters investigated include sperm motility via computer-assisted semen analysis (CASA), sperm concentration, and sperm biomarkers that promote paternal contribution of spermatozoa to fertilization including hyaluronic acid binding, chromatin maturity, apoptotic markers, cytoplasmic retention, and sperm DNA integrity.
As compared to TYM, NM was equally capable of sperm cryopreservation with both short-term and long-term storage in media, and after freeze-thaw and gradient processing of sperm. HA binding of sperm was comparable post thaw in both NM and yolk media. There are also no differences observed between the samples cryopreserved in NM or TYM in terms of their aniline blue staining, CK immunocytochemistry, caspase 3 immunostaining, or DNA nick translation.
NM has the advantage of being xeno-free, yet in preservation of sperm motility and other sperm attributes, the NM is as effective as the TYM.
我们旨在研发一种安全的精子冷冻保存新媒体(NM),其由不含任何动物源的一致且可重复的成分组成,并将NM与常用的卵黄培养基(TYM,欧文科学公司产品)相比,评估其对精子结构和功能的影响。
我们评估了患者精液样本,并将其分别在NM或TYM中一式两份进行冷冻保存。样本在解冻前分别短期冷冻保存1周或长期冷冻保存1个月。研究的参数包括通过计算机辅助精液分析(CASA)测定的精子活力、精子浓度,以及促进精子对受精的父系贡献的精子生物标志物,包括透明质酸结合、染色质成熟度、凋亡标志物、细胞质保留和精子DNA完整性。
与TYM相比,NM在短期和长期培养基保存中,以及在精子冻融和梯度处理后,均同样能够实现精子冷冻保存。解冻后,精子在NM和卵黄培养基中的透明质酸结合能力相当。在苯胺蓝染色、CK免疫细胞化学、半胱天冬酶3免疫染色或DNA缺口平移方面,NM或TYM冷冻保存的样本之间也未观察到差异。
NM具有无外源物质的优势,而且在保存精子活力和其他精子属性方面,NM与TYM同样有效。