Lorthongpanich Chanchao, Yang Shang-Hsun, Piotrowska-Nitsche Karolina, Parnpai Rangsun, Chan Anthony W S
Yerkes National Primate Research Center, Department of Human Genetics and Genetics and Molecular Biology Program, Emory University School of Medicine, North East Atlanta, Georgia 30329, USA.
Reproduction. 2008 Jun;135(6):805-13. doi: 10.1530/REP-07-0478.
The recently developed technique of establishing embryonic stem (ES) cell lines from single blastomeres (BTMs) of early mouse and human embryos has created significant interest in this source of ES cells. However, sister BTMs of an early embryo might not have equal competence for the development of different lineages or the derivation of ES cells. Therefore, single BTMs from two- and four-cell embryos of outbred mice were individually placed in sequential cultures to enhance the formation of the inner cell mass (ICM) and the establishment of embryonic outgrowth. The outgrowths were then used for the derivation of ES cell lines. Based on the expression of ICM (Sox2) and trophectoderm (Cdx2) markers, it was determined that ICM marker was lacking in blastocysts derived from 12% of BTMs from two-cell stage and 20% from four-cell stage. Four ES cell lines (5.6%; 4/72) were established ater culture of single BTMs from two-cell embryos, and their pluripotency was demonstrated by their differentiation into neuronal cell types. Our results demonstrate that sister BTMs of an early embryo are not equally competent for ICM marker expression. However, we demonstrated the feasibility of establishing ES cells from a single BTM of outbred mice.
最近从早期小鼠和人类胚胎的单个卵裂球（BTM）建立胚胎干细胞（ES）系的技术引起了人们对这种ES细胞来源的极大兴趣。然而，早期胚胎的姐妹卵裂球在不同谱系发育或ES细胞衍生方面可能没有同等能力。因此，将远交系小鼠二细胞和四细胞胚胎的单个卵裂球分别进行连续培养，以促进内细胞团（ICM）的形成和胚胎外植体的建立。然后将外植体用于ES细胞系的衍生。根据ICM（Sox2）和滋养外胚层（Cdx2）标志物的表达，确定来自二细胞期12%的BTM和四细胞期20%的BTM所形成的囊胚中缺乏ICM标志物。从二细胞胚胎的单个BTM培养后建立了4个ES细胞系（5.6%；4/72），并通过它们分化为神经元细胞类型证明了其多能性。我们的结果表明，早期胚胎的姐妹卵裂球在ICM标志物表达方面并非同等胜任。然而，我们证明了从远交系小鼠的单个BTM建立ES细胞的可行性。
