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.
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.
Genome Institute of Singapore, Singapore.
Petra
P
Kraus
Geraldine
G
Leong
Valerie
V
Tan
Xing
X
Xing
Jie Wei
JW
Goh
Sook Peng
SP
Yap
Thomas
T
Lufkin
D000818:Animals; D001755:Blastocyst; D002454:Cell Differentiation; D002678:Chimera / Q000235:genetics*; D003362:Cost-Benefit Analysis; D004622:Embryo, Mammalian / Q000166:cytology* / Q000502:physiology; D053595:Embryonic Stem Cells / Q000502:physiology*; D005260:Female; D005854:Germ Cells; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008845:Microinjections