Schiewe Mitchel C, Zozula Shane, Anderson Robert E, Fahy Gregory M
Ovagen Fertility/Southern California Institute for Reproductive Sciences (SCIRS), 361 Hospital Road, Suite 433, Newport Beach, CA 92663, USA.
Ovagen Fertility/Southern California Institute for Reproductive Sciences (SCIRS), 361 Hospital Road, Suite 433, Newport Beach, CA 92663, USA.
Cryobiology. 2015 Oct;71(2):264-72. doi: 10.1016/j.cryobiol.2015.07.009. Epub 2015 Jul 23.
A novel, aseptic closed system vitrification (VTF) technique for the cryopreservation of embryos and oocytes has been developed and clinically validated in this study. It combines the practicality of embryo-containing sterile flexipettes stored safely and securely with 0.3 ml CBS™ embryo straws possessing weld seals. The cooling and warming rates of this double container system were determined using a data logger. Upon direct plunging into LN(2), the flexipettes cool at an average rate of 1391°C/min, while warming occurs at an average rate of 6233°C/min in a 37°C 0.5 M sucrose bath. Direct deposition of the flexipette into a warming bath insured a rapid transition between -100 and -60°C to minimize potentially harmful recrystalization associated with devitrification. In conclusion, the μS-VTF system has exhibited higher (p<0.05) intact survival, implantation and live birth rates than conventional slow freezing methods. The effective embryo transfer of vitrified blastocysts proved similar to or better than fresh embryo transfer outcomes. The sustained clinical use of μS-VTF has justified a change in our infertility practice. Capsule: The microSecure vitrification (μS-VTF) procedure is a low-cost, non-commercial, aseptic, closed system that offers technical simplicity and repeatability, while effectively attaining an estimated 4:1 warming-to-cooling rate ratio, which supports excellent embryo survival and sustained viability.
本研究开发并临床验证了一种用于胚胎和卵母细胞冷冻保存的新型无菌封闭系统玻璃化冷冻(VTF)技术。它将安全储存的含胚胎无菌柔性移液管的实用性与具有焊接密封的0.3 ml CBS™胚胎吸管相结合。使用数据记录器测定了该双容器系统的冷却和升温速率。直接投入液氮后,柔性移液管在37°C的0.5 M蔗糖浴中的平均冷却速率为1391°C/分钟,而升温速率为6233°C/分钟。将柔性移液管直接放入温浴中可确保在-100至-60°C之间快速过渡,以尽量减少与玻璃化转变相关的潜在有害再结晶。总之,μS-VTF系统的完整存活率、着床率和活产率均高于传统慢速冷冻方法(p<0.05)。玻璃化囊胚的有效胚胎移植结果证明与新鲜胚胎移植相似或更好。μS-VTF的持续临床应用证明了我们不育治疗实践的改变。总结:微安全玻璃化冷冻(μS-VTF)程序是一种低成本、非商业化、无菌的封闭系统,技术简单且可重复,同时有效地实现了约4:1的升温和降温速率比,有利于胚胎的良好存活和持续活力。