The primary cell culture using cells from tail tip tissues were performed as described previously [22 , 23 (link)]. Briefly, the tail tip tissues from B6D2F1 mice (aged 8–10 weeks) and from large Japanese field mice were obtained from anesthetized animals. The cells were incubated
in DMEM medium supplemented with 10% fetal bovine serum and 2.5 µg/ml amphotericin B to allow for cell migration at 37°C under 5% CO2 in air. Cells undergoing migration
were used for passaged culture or stocked after freezing using a CELLBANKER 1 (Takara Bio, Shiga, Japan; CB011). As donor cells, the tail tip cells derived from laboratory mice
were used at passages 3 to 5, and the tail tip cells derived from large Japanese field mice were used at passages 8 to 14. The preparation of donor cells for SCNT or iSCNT were
performed as previously described [20 (link), 21 ]. Briefly, the cell pellets were re-suspended in Hepes-CZB
(HCZB) medium supplemented with 6% d-BSA and placed on ice until cell fusion.
in DMEM medium supplemented with 10% fetal bovine serum and 2.5 µg/ml amphotericin B to allow for cell migration at 37°C under 5% CO2 in air. Cells undergoing migration
were used for passaged culture or stocked after freezing using a CELLBANKER 1 (Takara Bio, Shiga, Japan; CB011). As donor cells, the tail tip cells derived from laboratory mice
were used at passages 3 to 5, and the tail tip cells derived from large Japanese field mice were used at passages 8 to 14. The preparation of donor cells for SCNT or iSCNT were
performed as previously described [20 (link), 21 ]. Briefly, the cell pellets were re-suspended in Hepes-CZB
(HCZB) medium supplemented with 6% d-BSA and placed on ice until cell fusion.
