Derivation and characterization of bovine embryonic stem cells from somatic cell nuclear transfer and in vitro fertilized bovine embryos

J. RODRIGUES SANGALLI; M NAVARRO; P.J ROSS

California

Conferencia; Transgenic Animal Research Conference (TARC) 2019; 2019

UC Davis

Efficient derivation and long-term culture of bovine embryonic stem cells (bESCs) from blastocyst stage embryos was recently reported. These cells were established in a custom- made mTeSR medium devoid of TGFß supplemented with FGF2 and IWR1, a WNT signaling inhibitor. We have now adapted the culture conditions to use a commercially available base medium (N2B27) and tested its capacity for isolation of bESCs from SCNT and parthenogenetic embryos (P A). Zona-free IVF, SCNT and P A blastocysts were plated over an irradiated mouse embryonic fibroblasts and cultured in N2B27 medium containing FGF2 and IWR1. The whole culture was passaged once a week onto a fresh feeder layer by single cell dissociation. After 4 weeks in culture, well defined colonies appeared in the three embryo groups; however, the cell line derivation efficiently was significantly higher (ANOVA, P