Establishment of feeder layer culture system supported development of bovine embryonic stem cells

The culture system of feeder layer on Bovine embryonic stem cells were established in DMEM culture medium supplemented with 0.1 μmol/L Na 2SeO 3,15% NBS and 0.1 mmol/L β mercaptoethanol(Sigma).The results showed that 5 passage murine embryonic fribroblasts (MEF) and 5 passage new bovine testicular fibroblasts (NBTF) were isolated and cloned;Comparing with MEF, the length and width of NBTF were biger, inddition to, NBTF cell growed fast; Murine embryos from 12d to 16d were most suitable for isolation and clone of MEF; Treated with 0.25%trypsin+0.04% EDTA, single MEF cell and single NBTF cell were isolated and cloned by short term (from 2 min to 3 min); To supplement with 0.1 μmol/L Na 2SeO 3, 15% NBS and 0.1 mmol/L β mercaptoethanol is beneficial to reproduction of MEF and NBTF cells.