Effects of vitrification cryopreservation on follicular morphology and stress relaxation behaviors of human ovarian tissues: sucrose versus trehalose as the non-permeable protective agent
Effects of vitrification cryopreservation on follicular morphology and stress relaxation behaviors of human ovarian tissues: sucrose versus trehalose as the non-permeable protective agent
复制标题
玻璃化冷冻保存对人卵巢组织卵泡形态和应力松弛行为的影响：蔗糖与海藻糖作为非渗透性保护剂
DOI：
10.1093/humrep/dev012
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发表时间：
2015
影响因子：
6.1
通讯作者：
Zhou Ping
中科院分区：
文献类型：
--
作者：
Tian Ting;Zhao Gang;Han Dan;Zhu Kaixuan;Chen Dawei;Zhang Zhiguo;Wei Zhaolian;Cao Yunxia;Zhou Ping
研究问题 蔗糖在人卵巢组织冷冻保存中比海藻糖更有效吗？ 总结答案 蔗糖作为冷冻保护剂（CPA）在人卵巢组织冷冻保存中的效果与海藻糖没有显着差异。已知的信息糖具有保持细胞膜完整的能力，并且可以降低 CPA 的毒性。蔗糖是最常用的非渗透性CPA，而海藻糖很少用于人卵巢组织冻存。尽管有多种方法来评估人类卵巢组织冷冻保存的效率，但很少有研究从生物力学的角度评估冷冻保存的效果。 研究设计、大小、持续时间从15名良性卵巢肿瘤或恶性肿瘤患者（20-41岁）中收集了总共15个卵巢组织样本，每个样本被切成六片。取两片作为新鲜对照组。其余四片切片使用不同的玻璃化方案进行玻璃化。升温后，将每组样品固定用于组织学评估或用于应力松弛测试。 参与者/材料、设置、方法对照组和玻璃化组的 CPA 溶液由 EDS 和 EDT（E，乙二醇；D，二甲亚砜；S，蔗糖；T，海藻糖）组成。使用动态机械分析仪在室温下进行应力松弛实验。评估卵巢组织样本的形态和粘弹性。应力松弛数据（SRD）以百分比计算，代表拉伸后保持初始应力的能力。比较各组间形态正常卵泡的百分比，以形态保存率表示。 主要结果及机会的作用新鲜对照组原始卵泡形态保存率（87.58%）高于S组（72.33%）（P=0.000）和T组（79.56%）（P=0.002）。尽管没有统计学意义，但与 S 组相比，T 组玻璃化冷冻表明原始卵泡的形态保存率有更高的趋势。新鲜对照组的SRD（0.6433±0.7233）与S组（0.5200±0.8331，P=0.000）和T组（0.5667±0.6415，P=0.000）差异有统计学意义。然而，S 组和 T 组之间没有发现显着差异。限制，注意原因实验样品直接暴露在空气中，这将导致实验组织和体内组织之间的粘弹性特性存在差异。研究结果的更广泛意义我们的研究表明，与蔗糖相比，海藻糖玻璃化后原始卵泡的形态保存率有更高的趋势，这可能提供为进一步优化人卵巢组织玻璃化冷冻奠定基础。此外，还可以从生物力学的角度评估卵巢组织冷冻保存的效果。 研究经费/竞争利益本研究得到了安徽省卫生部医学科学研究课题（2010B014）和国家基础研究计划（973计划）（2012CB944704）和国家自然科学基金（Nos. 51276179 和 51476160）。作者声明不存在冲突......
STUDY QUESTIONIs sucrose more effective than trehalose in human ovarian tissue cryopreservation?SUMMARY ANSWERThe effect of sucrose as a cryoprotective agent (CPA) was not significantly different from that of trehalose in human ovarian tissue cryopreservation.WHAT IS KNOWN ALREADYSugars have the ability to keep the cell membrane intact and can decrease the toxicity of CPAs. Sucrose is the most commonly used non-permeable CPA, while trehalose is rarely used in human ovarian tissue cryopreservation. Although various methods are utilized to evaluate the efficiency of human ovarian tissue cryopreservation, few studies have evaluated the effect of cryopreservation from the viewpoint of biomechanics.STUDY DESIGN, SIZE, DURATIONA total of 15 ovarian tissue samples were collected from 15 patients (20–41 years old) with benign ovarian tumors or malignancies, and each was dissected into six slices. Two slices were taken as the fresh control group. The remaining four slices were vitrified using different vitrification protocols. After warming, samples in each group were either fixed for histological evaluation or destined for stress relaxation test.PARTICIPANTS/MATERIALS, SETTING, METHODSThe CPA solutions for the control and vitrified groups were composed of EDS and EDT (E, ethylene glycol; D, dimethylsulphoxide; S, sucrose; T, trehalose). The stress relaxation experiments were carried out at room temperature using a dynamic mechanical analyzer. Ovarian tissue samples were assessed for both their morphology and viscoelasticity. Stress relaxation data (SRD) were calculated as a percentage, representing the ability to maintain the initial stress after stretching. The percentage of morphologically normal follicles was compared between groups, which was represented by morphologic preservation ratio.MAIN RESULTS AND THE ROLE OF CHANCEThe morphologic preservation ratio of the primordial follicles in the fresh control group (87.58%) was higher than that in group S (72.33%) (P= 0.000) and group T (79.56%) (P= 0.002). Although not statistically significant, compared with the S group, vitrification with T suggested a trend toward a higher morphologic preservation ratio of the primordial follicles. The SRD in the fresh control group (0.6433 ± 0.7233) was significantly different from that in group S (0.5200 ± 0.8331,P= 0.000) or in group T (0.5667 ± 0.6415,P= 0.000). However, no significant difference was found between groups S and T.LIMITATIONS, REASONS FOR CAUTIONExperimental samples were directly exposed to the air, which will result in a discrepancy in the viscoelastic properties between experimental tissues andin vivotissues.WIDER IMPLICATIONS OF THE FINDINGSOur study suggested a trend toward a higher morphologic preservation ratio of the primordial follicles after vitrification in trehalose compared with sucrose, which may provide a basis for further optimizing human ovarian tissue vitrification. In addition, it was possible to evaluate the effect of ovarian tissue cryopreservation from a biomechanics perspective.STUDY FUNDING/COMPETING INTEREST(S)This study was supported by the grants from the Medical Scientific Research Subject, Health Ministry of Anhui Province (2010B014) and National Basic Research Program of China (973 Program) (2012CB944704), and the National Natural Science Foundation of China (Nos. 51276179 and 51476160). The authors declare that there is no conflict of …