Mouse Embryonic stem cells (ESCs) were isolated from mouse (Strain C57BL⁄6) embryos at Day E3.5. Before cryopreservation, the cells were tested for viability (>70% post-thaw viability), their ability to remain undifferentiated when cultured on mouse feeder cells as assayed by marker expression (Oct4, SSEA-1, Nanog) and their ability to differentiate into embryoid bodies and express markers indicative of three germ layers (Ectoderm: (+) for β3-tubulin and nestin, Mesoderm: (+) for smooth muscle actin, Endoderm: (+) for AFP). Cells were cryopreserved at passage 20 (P20).

For optimal growth performance, we recommend culturing and expanding the cells in Knockout DMEM (SKU# 10829-018), Knockout serum replacement (SKU# 10828-028), Non-essential amino acids solution (SKU# 11140-050), L-glutamine (SKU# 25030081) and Recombinant human leukemia inhibitory factor (SKU# PHC9464).