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Ovarian tissue cryopreservation is an important method for female fertility preservation. Slow freezing of ovarian tissue results in poor follicular survival and low retransplantation efficiency. This study optimized the ovarian tissue cooling procedure by ice seeding, and the effects of ice seeding temperature and cooling rate after seeding on ovarian tissue cryopreservation were analyzed. The programmed cooling apparatus was combined with an ultrasonic device to achieve the ultrasonic seeding of ice crystals, and the ultrasonic intensity was screened. The ovarian survival and histology were assessed after rewarming. The results revealed that the optimized cooling procedure with ice seeding reduced the damage to ovarian tissues. When ice seeding was triggered at -11 ℃ with a cooling rate of 1 ℃/min after nucleation, follicle survival was 88.02%. Ultrasonic nucleation equipment enabled contactless ice seeding of the samples, reducing the risk of contamination and improving the success rate of ice seeding. Furthermore, the follicle survival rate of frozen ovarian tissue increased to 88.38%. The optimization of the procedure and the improvement of the equipment improved the effect of ovarian tissue cryopreservation, reduced the risk of introducing contamination during the cryopreservation process, and provided a new method for the slow cryopreservation of ovarian tissues in clinics.
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Zhou Xinli, female, professor, Institute of Biothermal Science, University of Shanghai for Science and Technology, 86-21-55270218, E-mail:
zjulily@163.com. Research fields: cryopreservation technology for cells, tissues and organs.
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卵巢组织冻存是女性生育力保存的重要方法,慢速冷冻卵巢组织卵泡存活率较低,自体移植效率低。通过诱发成核对卵巢组织降温程序进行优化,分析了成核温度和成核后降温速率对卵巢组织冷冻保存效果的影响。将程序降温仪与超声波诱发成核设备结合,实现超声波诱发成核并优化了超声波强度,对复温后的卵巢存活率和组织学形态进行评估。结果表明:引入诱发成核的降温程序对卵巢组织损伤减小,当在-11 ℃诱发成核,成核后降温速率为1 ℃/min时,卵泡存活率达88.02%。超声波成核设备可对样品进行无接触式诱发成核,降低了污染风险,提高了成核的成功率,冻存卵巢组织卵泡存活率达88.38%。通过程序的优化以及设备的改进提高了卵巢组织冷冻保存效果,降低了冷冻保存过程中引入污染的风险,为临床上卵巢组织慢速冷冻保存提供了新的方法。
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周新丽,女,教授,上海理工大学生物系统热科学研究所,021-55270218,E-mail:
zjulily@163.com。研究方向:细胞、组织、器官的低温保存技术。
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Ice seeding with precooled needle, figureFileSmall=YzP+y/D6Hc3f/a0zBDGL+A==, figureFileBig=TH7oeJK25tq8ayh2Wl1dwg==, tableContent=null), ArticleFig(id=1239230406785036639, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=CN, label=图1, caption=
预冷探针接触诱发成核示意图, figureFileSmall=YzP+y/D6Hc3f/a0zBDGL+A==, figureFileBig=TH7oeJK25tq8ayh2Wl1dwg==, tableContent=null), ArticleFig(id=1239230407032500597, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=EN, label=Fig.2, caption=
Design of ovarian tissue slow freezing procedure with ice seeding by precooled needle, figureFileSmall=8n+9av0vnagnLiXKWmrJbw==, figureFileBig=665BsiRCulaAYM8TivoH6w==, tableContent=null), ArticleFig(id=1239230407141552513, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=CN, label=图2, caption=
预冷探针诱发成核卵巢组织慢速冷冻程序设计, figureFileSmall=8n+9av0vnagnLiXKWmrJbw==, figureFileBig=665BsiRCulaAYM8TivoH6w==, tableContent=null), ArticleFig(id=1239230407275770251, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=EN, label=Fig.3, caption=
Ultrasonic ice seeding freezing device, figureFileSmall=r2eB/hkH6bKEDoYSkAECyQ==, figureFileBig=nKiQBgSNcJevOTuImIRPtg==, tableContent=null), ArticleFig(id=1239230407380627859, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=CN, label=图3, caption=
超声波诱发成核冻存设备, figureFileSmall=r2eB/hkH6bKEDoYSkAECyQ==, figureFileBig=nKiQBgSNcJevOTuImIRPtg==, tableContent=null), ArticleFig(id=1239230407464513945, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=EN, label=Fig.4, caption=
Survival rate of follicle with different cooling procedures under ice seeding by precooled needle, figureFileSmall=eHDNTClWK57EP5lN3V5anQ==, figureFileBig=XbFslym/n+NjupBrTy0lqQ==, tableContent=null), ArticleFig(id=1239230407577760158, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=CN, label=图4, caption=
预冷探针诱发成核下不同降温程序冻存卵巢组织卵泡存活率, figureFileSmall=eHDNTClWK57EP5lN3V5anQ==, figureFileBig=XbFslym/n+NjupBrTy0lqQ==, tableContent=null), ArticleFig(id=1239230409066738087, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=EN, label=Fig.5, caption=
HE of different cooling procedures under ice seeding by precooled needle, figureFileSmall=KSHwsT9Yg6NvJGZN8hlATw==, figureFileBig=5wkSqSfoWiUgGbp4K3UY7w==, tableContent=null), ArticleFig(id=1239230409167401391, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=CN, label=图5, caption=
预冷探针诱发成核冻存卵巢组织HE染色, figureFileSmall=KSHwsT9Yg6NvJGZN8hlATw==, figureFileBig=5wkSqSfoWiUgGbp4K3UY7w==, tableContent=null), ArticleFig(id=1239230409297424822, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=EN, label=Fig.6, caption=
Effect of ultrasonic ice seeding on the survival rate of follicle, figureFileSmall=wzblLwNhjAKwC4G68YxKIQ==, figureFileBig=2Q0iLw+zwsstzLDMdopoMg==, tableContent=null), ArticleFig(id=1239230409469391291, tenantId=1146029695717560320, journalId=1238823019242635269, articleId=1239230399931543586, language=CN, label=图6, caption=
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