Kraus Petra, Leong Geraldine, Tan Valerie, Xing Xing, Goh Jie Wei, Yap Sook Peng, Lufkin Thomas
Genome Institute of Singapore, Singapore.
Genesis. 2010 Jun;48(6):394-9. doi: 10.1002/dvg.20627.
The long-standing traditional method of delivering embryonic stem (ES) cells adjacent to the inner cell mass (ICM) of blastocysts to generate chimeras improved with the advent of laser- or Piezo assisted 8-cell embryo microinjection. Building on this technology but omitting either the laser or the Piezo to penetrate the zona pellucida and making use of earlier embryonic stages (2-cell and 4-cell), we were able to significantly speed up and economize our ES cell microinjection and chimera production throughput. We demonstrate here that embryonic (ES) and induced pluripotent stem (iPS) cells can stay fully pluripotent when delivered into 2-cell- and 4-cell-stage embryos, long before they would naturally be incorporated into the ICM of a blastocyst (E3.5) and give rise to high percentage and germline transmitting chimeras.
随着激光或压电辅助8细胞胚胎显微注射技术的出现，将胚胎干细胞（ES）细胞输送到囊胚内细胞团（ICM）附近以产生嵌合体的长期传统方法得到了改进。基于这项技术，但省略了激光或压电穿透透明带，并利用更早的胚胎阶段（2细胞和4细胞），我们能够显著加快并节省ES细胞显微注射和嵌合体生产的通量。我们在此证明，当将胚胎（ES）和诱导多能干细胞（iPS）细胞输送到2细胞和4细胞期胚胎中时，它们可以保持完全多能性，这远远早于它们自然地整合到囊胚的ICM（E3.5）中并产生高比例且能进行种系传递的嵌合体的时间。
