Culture conditions have been established for a second blastocyst-derived cell line, trophoblast stem (TS) cells, in addition to embryonic stem (ES) cells. This protocol describes a method for culturing TS cell lines. These cells can then be used to study trophoblast differentiation and placental function. - [Read Culturing Trophoblast Stem (TS) Cell Lines Protocol]
Protocol describes a method for removal of mouse ovaries in combination with the administration of progesterone, which causes the arrest of blastocyst development. This procedure is performed during the afternoon of the third day of pregnancy to ensure that the morulae have moved sufficiently into the oviducts and will be less likely to be damaged during the ovariectomy. - [Read Ovariectomy for Induction of Blastocyst Implantation Delay Protocol]
Details a placenta specific gene manipulation by transducing blastocysts with lentiviral vectors1. After a removal of zona pellucida which functions as a physical barrier, trophoblast cells lying outermost layer of blastocyst were transduced from outside with high-titer lentiviral vectors. As most placental cells descend from trophoblast cells while fetus originated from inner cell mass, transgene expression can be observed in trophoblast cells from preimplantation stages and in placenta... - [Read Placenta Specific Gene Manipulation by Transducing Zona-Free Blastocyst using Lentiviral Vector]
Blastocyst transfer is usually performed 24 hours after aggregation, when the morulae have become expanded blastocysts, and on the same day as injection. A little time is given between injection and transfer to allow the blastocysts to re-expand. Includes: The Mouse Recipient; The Transfer. - [Read Transfer of Injected Blastocysts to Pseudo Pregnant Mice Protocol]
The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.