Modifications made to culture medium by bovine oviduct epithelial cells: Changes to carbohydrates stimulate bovine embryo development

摘要: Co-culture remains a common method to support the development of bovine embryos, derived from IVM/IVF procedures. However, mechanism by which somatic cells confer their benefit developing embryo undetermined. This study therefore analysed changes made culture medium TCM-199, used in co-culture systems, and determined effects specific composition on culture. Bovine oviduct epithelial (BOE), Buffalo rat liver (BRL) fibroblast (3T3) were compared. The concentrations glucose, L-lactate, pyruvate, amino acids, NH4+, H+ gas tensions O2 CO2 measured TCM-199 supplemented with 10% fetal calf serum (FCS) prior directly following 48 h incubation periods each cell type. All three types modified carbohydrate media similar manner greatest BOE cells. Notable alterations an increase levels L-lactate pyruvate reduction glucose concentration, case cells, fell 5.55 mM 2.67 mM. In order determine relevance such development, modifications synthetic fluid (SOF) effect blastocyst vitro assessed. SOF acids BSA (SOFaa), significantly more zygotes developed stage (64%; P < 0.01) than SOFaa D/L-lactate equivalent those (11%). Interestingly, when carbohydrates mimicked present 57% reached stage. improvement was ascribed increases system. Results this demonstrate that create environment favourable embryonic development. analysis samples enzymatic methods meant only biologically active L-isomer lactate quantified. SOFaa, both inactive D-isomer are equimolar amounts. As such, syrup is actually contain 50% meaning all reported at twice effective concentration. Therefore concentration subsequently study. Blastocyst poor (24-36%) until total system equal or greater 0.82 numbers remained low (60.1 +/- 6.9 - 78.5 6.6) 3.3 data reinforces morphological appearance not sensitive enough be as sole criterion for assessing