Cryopreservation of germline stem cells in American paddlefish (Polyodon spathula)

文献类型： 外文期刊

第一作者： Ye, Huan

作者： Ye, Huan;Zhou, Congli;Yue, Huamei;Wu, Mengbin;Ruan, Rui;Du, Hao;Li, Chuangju;Wei, Qiwei

作者机构：

关键词： Cryopreservation; Germline stem cells; Acipenseriformes; Germ cell transplantation

期刊名称：ANIMAL REPRODUCTION SCIENCE （ 影响因子：2.145； 五年影响因子：2.281 ）

ISSN： 0378-4320

年卷期： 2021 年 224 卷

页码：

收录情况： SCI

摘要： Most sturgeon and paddlefish are critically endangered; therefore, effective measures to conserve these genetic resources are required. Cryopreservation of gonad tissues containing germline stem cells could be an effective strategy for long term preservation and restoration of fish species using germ cell transplantation procedure. The aim of this study was to develop an optimal procedure for long-term cryopreservation of American paddlefish gonads using a slow-freezing method. Through optimization of permeating cryopmtectants, nonpermeating cryoprotectants, and supplementation of proteins, gonad tissues were frozen with a cryomedium containing 1.3 M dimethyl sulfoxide, 0.1 M trehalose, and 10 % fetal bovine serum at a cooling rate of -1 degrees C/min. This method was also successfully utilized for the cryopreservation of Yangtze sturgeon testes. Viability of gonadal cells isolated from frozen gonads was not different from cells isolated from fresh gonadal tissues, while the number of gonadal cells dissociated from frozen gonads was less. Germline stem cells dissociated from long-term (1 year) cryopreserved gonads were labeled with PKH26 fluorescent dye and intraperitoneally transplanted into larvae of Yangtze sturgeon. The colonization of transplanted germline stem cells was confirmed by the presence of PKH26-labeled donor germline stem cells and donor-derived mtDNA sequence in the recipient gonads, providing evidence that germline stem cells from sturgeon and paddlefish gonads that had been preserved for a long period maintained their functions. The results of present study indicate the procedures used are effective for long-term preservation of critically endangered species within the Acipenseriformes order which can later be regenerated using surrogate broodstock technology.

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