Efficient establishment of mouse embryonic stem cell lines from single Blastomeres and polar bodies
Efficient establishment of mouse embryonic stem cell lines from single Blastomeres and polar bodies
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DOI：
10.1634/stemcells.2006-0615
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发表时间：
2007-01-01
期刊：
影响因子：
5.2
通讯作者：
Wakayama, Teruhiko
中科院分区：
文献类型：
--
作者：
Wakayama, Sayaka;Hikichi, Takafusa;Wakayama, Teruhiko
最近，从小鼠和人类的8细胞胚胎中提取的单个卵裂球建立了ES细胞系，成功率分别为4%和2%，这表明该方法可以用于再生医学，以减少对胚胎伤害的伦理担忧。然而，这些研究使用了其他ES细胞作为支持细胞。在这里，我们报道了一种从单个卵裂球建立小鼠ES细胞系的简单而高效的方法，在这种方法中，单个卵裂球只需用改良的ES细胞培养液移植到小鼠胚胎成纤维细胞的饲养层上。从2细胞胚胎(发育率为50%~69%)、早期4细胞胚胎(28%~40%)、4细胞晚期胚胎(22%)和8细胞胚胎(14%~16%)共建立了112个ES细胞系。我们还成功地从第一极体(36%-40%)和第二极体(33%)建立了18个孤雌生殖ES细胞系，其核通过核移植重建为胚胎。大多数被检查的细胞系保持了正常的核型，并表达了多能性的标记，包括在嵌合体小鼠中的生殖系传播。我们的结果表明，所有早期胚胎或极体的单个细胞都有潜力转化为ES细胞，而不需要任何特殊处理。
Recently, ES cell lines were established from single blastomeres taken from eight-cell embryos in mice and humans with success rates of 4% and 2%, respectively, which suggests that the method could be used in regenerative medicine to reduce ethical concerns over harm to embryos. However, those studies used other ES cells as supporting cells. Here, we report a simple and highly efficient method of establishing mouse ES cell lines from single blastomeres, in which single blastomeres are simply plated onto a feeder layer of mouse embryonic fibroblasts with modified ES cell medium. A total of 112 ES cell lines were established from two-cell (establishment rate, 50%-69%), early four-cell (28%-40%), late four-cell (22%), and eight-cell (14%-16%) stage embryos. We also successfully established 18 parthenogenetic ES cell lines from first (36%-40%) and second polar bodies (33%), the nuclei of which were reconstructed to embryos by nuclear transfer. Most cell lines examined maintained normal karyotypes and expressed markers of pluripotency, including germline transmission in chimeric mice. Our results suggest that the single cells of all early-stage embryos or polar bodies have the potential to be converted into ES cells without any special treatment.