A More Cost Effective and Rapid High Percentage Germ-Line Transmitting Chimeric Mouse Generation Procedure via Microinjection of 2-Cell, 4-Cell, and 8-Cell Embryos With ES and iPS Cells

作者：Kraus Petra; Leong Geraldine; Tan Valerie; Xing Xing; Goh Jie Wei; Yap Sook Peng; Lufkin Thomas^*

来源：Genesis, 2010, 48(6): 394-399.

DOI：10.1002/dvg.20627

• ES cell microinjection
• 8C embryo
• 4C embryo
• 2C embryo
• germline transmission
• nonlaser assisted

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. genesis 48: 394-399, 2010.

• 出版日期2010-6