Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, No. 800. Dongchuan Road, Minhang District, Shanghai 200240, China.
Mol Hum Reprod. 2014 Mar;20(3):271-81. doi: 10.1093/molehr/gat081. Epub 2013 Nov 20.
Germline stem cell lines possess the abilities of self-renewal and differentiation, and have been established from both mouse and human ovaries. Here, we established a new female germline stem cell (FGSC) line from post-natal rats by immunomagnetic sorting for Fragilis, which showed a normal karyotype, high telomerase activity, and a consistent gene expression pattern of primordial germ cells after 1 year of culture. Using an in vitro differentiation system, the FGSC line could differentiate into oocytes. After liposome-based transfection with green fluorescent protein (GFP) or fat-1 vectors, the FGSCs were transplanted into the ovaries of infertile rats. The transplanted FGSCs underwent oogenesis, and the rats produced offspring carrying the GFP or fat-1 transgene after mating with wild-type male rats. The efficiency of gene transfer was 27.86-28.00%, and 2 months was needed to produce transgenic rats. These findings have implications in biomedical research and potential applications in biotechnology.
成体生殖干细胞系具有自我更新和分化的能力，已从人和鼠的卵巢中建立。在这里，我们通过免疫磁珠分选 Fragilis 建立了一个新的来自于出生后大鼠的雌性生殖干细胞系（FGSC），其表现出正常的核型、高端粒酶活性，并且在培养 1 年后保持原始生殖细胞的一致基因表达模式。使用体外分化系统，FGSC 系可以分化为卵母细胞。在用脂质体转染 GFP 或 fat-1 载体后，FGSCs 被移植到不孕大鼠的卵巢中。移植的 FGSCs 经历了卵发生，并且这些大鼠在与野生型雄性大鼠交配后产生了携带 GFP 或 fat-1 转基因的后代。基因转移的效率为 27.86-28.00%，需要 2 个月的时间才能产生转基因大鼠。这些发现对生物医学研究具有重要意义，并可能在生物技术中有潜在的应用。
