Müller Thomas, Fleischmann Gesine, Eildermann Katja, Mätz-Rensing Kerstin, Horn Peter A, Sasaki Erika, Behr Rüdiger
Stem Cell Research Group, German Primate Center, Kellnerweg 4, 37077 Göttingen, Germany.
Hum Reprod. 2009 Jun;24(6):1359-72. doi: 10.1093/humrep/dep012. Epub 2009 Feb 27.
Embryonic stem cells (ESC) hold great promise for the treatment of degenerative diseases. However, before clinical application of ESC in cell replacement therapy can be achieved, the safety and feasibility must be extensively tested in animal models. The common marmoset monkey (Callithrix jacchus) is a useful preclinical non-human primate model due to its physiological similarities to human. Yet, few marmoset ESC lines exist and differences in their developmental potential remain unclear.
Blastocysts were collected and immunosurgery was performed. cjes001 cells were tested for euploidy by karyotyping. The presence of markers for pluripotency was confirmed by immunofluorescence staining and RT-PCR. Histology of teratoma, in vitro differentiation and embryoid body formation revealed the differentiation potential.
cjes001 cells displayed a normal 46,XX karyotype. Alkaline phosphatase activity, expression of telomerase and the transcription factors OCT4, NANOG and SOX2 as well as the presence of stage-specific embryonic antigen (SSEA)-3, SSEA-4, tumor rejection antigens (TRA)-1-60, and TRA-1-81 indicated pluripotency. Teratoma formation assay displayed derivatives of all three embryonic germ layers. Upon non-directed differentiation, the cells expressed the germ cell markers VASA, BOULE, germ cell nuclear factor and synaptonemal complex protein 3 and showed co-localization of VASA protein within individual cells with the germ line stem cell markers CD9, CD49f, SSEA-4 and protein gene product 9.5, respectively.
The cjes001 cells represent a new pluripotent ESC line with evidence for enhanced spontaneous differentiation potential into germ cells. This cjes001 line will be very valuable for comparative studies on primate ESC biology.
胚胎干细胞（ESC）在治疗退行性疾病方面具有巨大潜力。然而，在ESC能够用于细胞替代疗法的临床应用之前，其安全性和可行性必须在动物模型中进行广泛测试。普通狨猴（Callithrix jacchus）由于其生理特征与人类相似，是一种有用的临床前非人类灵长类动物模型。然而，狨猴ESC系很少，其发育潜能的差异仍不清楚。
收集囊胚并进行免疫手术。通过核型分析检测cjes001细胞的整倍性。通过免疫荧光染色和逆转录聚合酶链反应（RT-PCR）确认多能性标志物的存在。畸胎瘤组织学、体外分化和胚状体形成揭示了分化潜能。
cjes001细胞显示正常的46,XX核型。碱性磷酸酶活性、端粒酶表达以及转录因子OCT4、NANOG和SOX2的表达，以及阶段特异性胚胎抗原（SSEA）-3、SSEA-4、肿瘤排斥抗原（TRA）-1-60和TRA-1-81的存在表明其具有多能性。畸胎瘤形成试验显示了所有三个胚胎胚层的衍生物。在非定向分化时，细胞表达生殖细胞标志物VASA、BOULE、生殖细胞核因子和联会复合体蛋白3，并显示VASA蛋白在单个细胞内分别与生殖系干细胞标志物CD9、CD49f、SSEA-4和蛋白基因产物9.5共定位。
cjes001细胞代表一种新的多能ESC系，有证据表明其向生殖细胞自发分化的潜能增强。该cjes001系对于灵长类ESC生物学的比较研究将非常有价值。
