This line was isolated from blastocysts that had been frozen on day six of in vitro development after in vitro fertilization. Karyotype analysis revealed that the line was female [46, XX], and had a normal karyotype.
These cells can be maintained on MEFs, by manual dissection, or transferred to and maintained on Matrigel-coated plates in MEF-conditioned medium. According to the provider's protocol, the growth medium should contains : DMEM (75%) supplemented with KSR (20%), GlutaMAX (2mM), MEM-NEAA (0.1mM), 2-mercaptoethanol (2mM), B27 supplement (without vitamin A), pen-strep (x1) and bFGF (20ng/ml).