I used a pastiche of three original sources:
- Manipulating the Mouse embryo, 3 ed. Protocol 4.11: "Collecting Blastocysts" was employed for isolation of 3.5dpc blastocysts.
- Manipulating the Mouse Embryo, 3 ed. Protocol 8.5: "De novo isolation of embryonic stem cells from blastocysts" was employed for disaggregating the embryo explants. (The most difficult part of the procedure)
- Bryja, V. et al. "An efficient method for derivation of mouse embryonic stem cells", Stem Cells 2006; 24:844-849, was employed for all the media formulations and timing of the disaggregation steps.