Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates.
J Vis Exp
; (135)2018 05 29.
Article
in En
| MEDLINE
| ID: mdl-29912196
ABSTRACT
In an effort to increase efficiency in the creation of genetically modified mice via ES Cell methodologies, we present an adaptation to the current blastocyst injection protocol. Here we report that a simple rotation of the embryo, and injection through Trans-Inner cell mass (TICM) increased the percentage of chimeric mice from 31% to 50%, with no additional equipment or further specialized training. 26 different inbred clones, and 35 total clones were injected over a period of 9 months. There was no significant difference in either pregnancy rate or recovery rate of embryos between traditional injection techniques and TICM. Therefore, without any major alteration in the injection process and a simple positioning of the blastocyst and injecting through the ICM, releasing the ES cells into the blastocoel cavity can potentially improve the quantity of chimeric production and subsequent germline transmission.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Chimerism
/
Embryonic Stem Cells
/
Microinjections
Limits:
Animals
/
Pregnancy
Language:
En
Journal:
J Vis Exp
Year:
2018
Document type:
Article