Assessment of immunoregulation by cultured, pre-attachment bovine embryos

The possibility that the viability of bovine embryos might be predicted by measuring their release of immunoregulatory substances during culture has been investigated. Bovine embryos between days 2 and 19 of gestation were cultured for 24-48 h, the embryo-conditioned medium was harvested and studied for suppression of PHA-stimulated bovine leukocyte cultures. Medium incubated in the absence of any conditioning tissues served as control. Artefactual immunosuppression was detected in incubated control material that could be attributed, in part, to the mixing of different tissue culture media, the type of plastic-ware employed for incubation and supplementation of media with additional L-glutamine. It was observed that day-2 to day-9 bovine embryos, cultured in medium able to support the lymphocyte proliferation assay, did not release immunosuppressive substances. Medium conditioned by day-10 to day-12 embryos produced variable immunosuppression while that conditioned by trophoblastic vesicles derived from day-14 to day-19 embryos was consistently highly suppressive. Since bovine embryo transfer is normally conducted at 6-8 days of gestation, it is unlikely that measuring the immunosuppressive products released from bovine embryos will be of value for predicting their viability.