This line was isolated from blastocysts that had been frozen at day six of in vitro development after in vitro fertilization. Karyotype analysis revealed that the line was male [46, XY], and had a normal karyotype. Specific molecular markers were detected, such as: SSEA4, Tra1-60 and alkaline phosphatase.
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).