In vitro co-culture of bovine embryos using Buffalo Rat Liver Cells, CR1aa and TMC199

Abstract

Seventeen hundred sixty-one in vitro produced bovine potential 1-cell embryos were divided into 12 treatment groups. Embryos were co-cultured with Buffalo Rat Liver Cells (BRLC) in CRlaa, without BRLC in CRlaa or with BRLC in Tissue Culture Medium 199 (TCM 1 99). The media were supplemented with polyvinyl alcohol (PVA), fetal calf serum (FCS), bovine serum albumin fraction V (BS"), or FCS + BS", respectively. In Experiment 1, the embryos were cultured at 3 70 C in a humidified atmosphere of 5% C02 in air. After 7 days, the number of oocytes developing to the CM and/or blastocyst stage was compared between different treatment groups. More embryos cultured in CRlaa with BRLC developed to the CM or blastocyst stage (P<.OS) when compared to embryos cultured in CRlaa alone or with BRLC in TCM199. Embryonic development was enhanced when FCS or FCS + BSAV was used in conjunction with CRlaa with BRLC, compared to TCM199 with BRLC., More embryos developed to the CM or blastocyst stage in CRlaa with BRLC than in CRlaa without BRLC (P<=.OS). In Experiment 2, compact morulae and blastocysts from Experiment I were cryopresevered, thawed and co-cultured for an additional 48 h to determine embryo viability by hatching. More embryos cultured in CRlaa with BRLC survived cryopreservation, than did those cultured in CRlaa without BRLC (P<.05) or in TCM199 with BRLC (P=.0632). No embryos cultured in CRlaa without BRLC hatched after cryopreservation. These results indicate that embryos can be cultured in a defined medium; however, no embryos cultured in the absence of BRLC survived cryopreservation.