Production of the medaka derived from vitrified whole testes by germ cell transplantation.
Production of the medaka derived from vitrified whole testes by germ cell transplantation.
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DOI：
10.1038/srep43185
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发表时间：
2017-03-03
影响因子：
4.6
通讯作者：
Yoshizaki G
中科院分区：
文献类型：
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作者：
Seki S;Kusano K;Lee S;Iwasaki Y;Yagisawa M;Ishida M;Hiratsuka T;Sasado T;Naruse K;Yoshizaki G
青鳉（Oryzias latipes）是一种硬骨鱼模式动物，与其他模式生物的区别在于存在近交系、野生种群和相关物种。低温保存保证了这些独特的生物资源的保存。然而，由于它们的体积大，它们的卵子和胚胎的冷冻保存技术尚未建立。在本研究中，我们建立了一种从冷冻保存的睾丸细胞（TC）中产生功能配子的方法。采用玻璃化法对青鳉的整个睾丸进行冷冻保存。解冻后，将从冷冻保存的睾丸组织中分离的细胞腹膜内移植到无菌的三倍体孵化的幼鱼中。一些细胞，可能是精原干细胞，迁移到受体的生殖嵴，并根据受体的性别产生卵子或精子。受体的交配导致成功生产冷冻保存的TC衍生后代。我们成功地从东京濒危野生种群加贺近交系中产生了个体，并从冷冻保存的TC中产生了亚生育突变体（wnt 4 b −/−）。这种方法有利于各种青鳉菌株的半永久保存。
The medaka (Oryzias latipes) is a teleost model distinguished from other model organisms by the presence of inbred strains, wild stocks, and related species. Cryopreservation guarantees preservation of these unique biological resources. However, because of their large size, cryopreservation techniques for their eggs and embryos have not been established. In the present study, we established a methodology to produce functional gametes from cryopreserved testicular cells (TCs). Whole testes taken from medaka were cryopreserved by vitrification. After thawing, the cells dissociated from cryopreserved testicular tissues were intraperitoneally transplanted into sterile triploid hatchlings. Some cells, presumably spermatogonial stem cells, migrated into the genital ridges of recipients and resulted in the production of eggs or sperm, based on sex of the recipient. Mating of recipients resulted in successful production of cryopreserved TC-derived offspring. We successfully produced individuals from the Kaga inbred line, an endangered wild population in Tokyo, and a sub-fertile mutant (wnt4b−/−) from cryopreserved their TCs. This methodology facilitates semi-permanent preservation of various medaka strains.