Long-term propagation of tree shrew spermatogonial stem cells in culture and successful generation of transgenic offspring
文献类型：期刊论文
| 作者 | Li CH1,2,3; Yan LZ4,5; Wang L1,2; Bi R4; Ji S1; Ma YH5; Nie WH1; Lv LB5; Ban WZ1; Yao YG[*]3,4,5,6 |
| 刊名 | CELL RESEARCH
 |
| 出版日期 | 2016 |
| 卷号 | 27期号:X页码:241-252 |
| 通讯作者 | zhaoxudong@mail.kiz.ac.cn ; zhengp@mail.kiz.ac.cn ; yaoyg@mail.kiz.ac.cn |
| 合作状况 | 其它 |
| 英文摘要 | Tree shrews have a close relationship to primates and have many advantages over rodents in biomedical research. However, the lack of gene manipulation methods has hindered the wider use of this animal. Spermatogonial stem cells (SSCs) have been successfully expanded in culture to permit sophisticated gene editing in the mouse and rat. Here, we describe a culture system for the long-term expansion of tree shrew SSCs without the loss of stem cell properties. In our study, thymus cell antigen 1 was used to enrich tree shrew SSCs. RNA-sequencing analysis revealed that the Wnt/β-catenin signaling pathway was active in undifferentiated SSCs, but was downregulated upon the initiation of SSC differentiation. Exposure of tree shrew primary SSCs to recombinant Wnt3a protein during the initial passages of culture enhanced the survival of SSCs. Use of tree shrew Sertoli cells, but not mouse embryonic fibroblasts, as feeder was found to be necessary for tree shrew SSC proliferation, leading to a robust cell expansion and long-term culture. The expanded tree shrew SSCs were transfected with enhanced green fluorescent protein (EGFP)-expressing lentiviral vectors. After transplantation into sterilized adult male tree shrew's testes, the EGFP-tagged SSCs were able to restore spermatogenesis and successfully generate transgenic offspring. Moreover, these SSCs were suitable for the CRISPR/Cas9-mediated gene modification. The development of a culture system to expand tree shrew SSCs in combination with a gene editing approach paves the way for precise genome manipulation using the tree shrew. |
| 收录类别 | SCI |
| 资助信息 | This work was supported by the Breakthrough Project of the Strategic Priority Program of the Chinese Academy of Scienc- es (Grant Nos XDB13000000 and XDA01010203) and the project from Yunnan Province to Ping Zheng (2015HA038). |
| 语种 | 英语 |
| 源URL | [http://159.226.149.26:8080/handle/152453/10751]  |
| 专题 | 昆明动物研究所_保护遗传学(细胞库) 昆明动物研究所_动物模型与人类重大疾病机理重点实验室 昆明动物研究所_遗传资源与进化国家重点实验室 昆明动物研究所_哺乳动物胚胎发育 昆明动物研究所_重大疾病机理的遗传学 昆明动物研究所_中国科学院昆明灵长类研究中心 |
| 作者单位 | 1.State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China 2.Yunnan Key Laboratory of Animal Reproduction, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China 3.Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming 650204, China 4.Key Laboratory of Animal Models and Human Disease Mechanisms, Kunming Institute of Zoology, Chinese Academy of Scienc- es, Kunming 650223, China 5.Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China 6.CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China |
| 推荐引用方式 GB/T 7714 | Li CH,Yan LZ,Wang L,et al. Long-term propagation of tree shrew spermatogonial stem cells in culture and successful generation of transgenic offspring[J]. CELL RESEARCH,2016,27(X):241-252. |
| APA | Li CH.,Yan LZ.,Wang L.,Bi R.,Ji S.,...&Wu Y.(2016).Long-term propagation of tree shrew spermatogonial stem cells in culture and successful generation of transgenic offspring.CELL RESEARCH,27(X),241-252. |
| MLA | Li CH,et al."Long-term propagation of tree shrew spermatogonial stem cells in culture and successful generation of transgenic offspring".CELL RESEARCH 27.X(2016):241-252. |
入库方式： OAI收割
来源：昆明动物研究所
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