Abstract
In this study we evaluated the use of a chemically defined medium in the production of blastocysts from bovine oocytes fertilized in vitro. As culture medium we used CRI-PVP, a modification of CRlaa medium with bovine serum albumin replaced by polyvinylpyrrolidone. After 168 h of culture (192 h after insemination) 8.7%, 10.5 and 12.8% of the cleaved embryos developed to the blastocyst stage in the presence of 0, 2 or 200 nM insulin, respectively. The supplementation of 200 nM insulin tended to increase cell numbers in morulae and blastocysts (P=0.10). It is concluded that CRI-PVP can be used as a chemically defined medium in the production of blastocysts from bovine 1-cell embryos. However, further modifications are needed, and the insulin concentrations used may be below the optimum for blastocyst production.
Highlights
The in vitro production (IVP) of bovine embryos is most efficiently carried out using serum- supplemented culture media and with co-culture with somatic cells
The effects of insulin on mouse embryo development have been characterized reasonably well (Rosenblum et al 1986, Harvey and Kaye 1988, 1991, Rao et al 1990), but relatively little is known about its effect on bovine embryos
The objective of our study was to evaluate the use of a chemically defined bovine embryo culture medium with polyvinylpyrrolidone (PVP) as a macromolecular substitute for serum or serum albumin. Using this medium, we examined the effect of insulin supplementation on bovine embryo development to the morula and blastocyst stage
Summary
The in vitro production (IVP) of bovine embryos is most efficiently carried out using serum- supplemented culture media and with co-culture with somatic cells (for review, see Brackett and Zuelke 1993). As serum consists of numerous unknown ingredients these media are termed undefined media Improving such a medium by analysing the supplementation of various ingredients is complicated as the medium may already include these ingredients. PDGF-oc are detectable throughout bovine preimplantation development (Watson et al 1992), implying that the use of these growth factors in culture media could have beneficial effects on embryonic development. The objective of our study was to evaluate the use of a chemically defined bovine embryo culture medium with polyvinylpyrrolidone (PVP) as a macromolecular substitute for serum or serum albumin. Using this medium, we examined the effect of insulin supplementation on bovine embryo development to the morula and blastocyst stage. After 24-h fertilization at 39°C in 5% C02 in air, the cumulus cells were removed by vortexing
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