Cryopreservation of human ovarian tissue by direct plunging into liquid nitrogen
Cryopreservation of human ovarian tissue by direct plunging into liquid nitrogen
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DOI：
10.1016/s0301-2115(02)00465-7
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发表时间：
2003-06-10
影响因子：
2.6
通讯作者：
Nawroth, F
中科院分区：
文献类型：
--
作者：
Isachenko, E;Isachenko, V;Nawroth, F
目的：为卵巢组织直接投入液氮冷冻保存方案的进一步发展确定一个有前景的方向。材料与方法：将20名患者的卵巢活检组织（切成3片0.5 mm厚的切片）暴露于：40%乙二醇+0.35 M蔗糖+5%蛋黄; 40%乙二醇+18% Ficoll-70 +0.35 M蔗糖; 20%乙二醇+20%二甲基亚砜。通过将0.25 ml吸管或铜网格插入液氮中或将0.25 ml吸管插入预冷（-196 ℃）的金属粉末中来完成碎片的冷冻保存。将解冻的碎片转移到蔗糖溶液中，以增量稀释冷冻保护剂。冻存后对组织进行组织学观察，并进行体外培养，研究冻存后激素产生能力。结果：只有在乙二醇-蔗糖-蛋黄液中超速冷却才能保护卵泡和间质免受损伤。结论：根据人卵巢组织直接投入液氮冷冻保存方案的要求，建立了以下参数：玻璃化冷冻液应包括乙二醇、二糖和蛋黄;超快速冷却/解冻应使用标准0.25吸管或铜网格。(C)2003年由Elsevier Science爱尔兰有限公司出版。
Aim: To establish a prospective direction for further development of the protocol for cryopreservation of ovarian tissue by direct plunging into liquid nitrogen. Materials and methods: Human ovarian biopsies from 20 patients (cut in similar to0.5 mm(3) pieces) were exposed to: 40% ethylene glycol + 0.35 M sucrose + 5% egg yolk; 40% ethylene glycol + 18% Ficoll-70 + 0.35 M sucrose; 20% ethylene glycol + 20% dimethyl sulphoxide. Cryopreservation of pieces was accomplished by plunging 0.25 ml straws or copper grids into liquid nitrogen or 0.25 ml straws into precooled (-196degreesC) metallic powder. Thawed pieces were transferred to sucrose solution for incremental dilution of cryoprotectants. Histological observation of the tissue was performed after cryopreservation and in vitro culture was done to study hormone production ability after cryopreservation. Results: Only ultrarapid cooling in ethylene glycol-sucrose-egg yolk solution protected both follicles and stroma from damage. Conclusion: The following parameters were established as required for a protocol of human ovarian tissue cryopreservation by direct plunging into liquid nitrogen: the vitrification medium should include ethylene glycol, disaccharide and egg yolk; ultrarapid cooling/thawing should take place using standard 0.25 straws or copper grids. (C) 2003 Published by Elsevier Science Ireland Ltd.