Persistence of the developmental block of in vitro fertilized domestic cat embryos to temporal variations in culture conditions

A series of studies examined the influence and temporal interaction of energy substrate, media complexity, and tissue co-culture on the development of in vitro fertilized cat embryos and the persistence of the morula-to-blastocyst developmental block. In Study I, oocytes were fertilized and cultured for 144 hr in a simple culture medium (modified Krebs Ringer bicarbonate; mKrb), containing either glucose or glutamine, or cultured in mKrb w/ glutamine for the initial 72 hr with transfer to mKrb w/ glucose for the final 72 hr. Fertilization rate, percent development to morulae, and cell number per embryo were similar (P > 0.05) between treatments and blastocyst formation was universally low (<10%). In Study II, oocytes were fertilized and cultured in either mKrb (w/ glucose or glutamine) or in a complex medium, Ham's F10 (w/ 10% fetal bovine serum [FBS]). After 72 hr of initial culture, embryos in mKrb were transferred into Ham's F10. Fertilization rate was lower (P < 0.01) in Ham's F10 but embryo development to the morulae stage and cell number per embryo were comparable (P > 0.05) for all treatments. A higher percentage of blastocysts and morulae becoming blastocysts were observed after initial culture in mKrb w/ glutamine than after initial culture in mKrb w/ glucose. In Study III, oocytes were fertilized and cultured initially in mKrb (w/ glutamine), and then switched to either Ham's F10 or cat oviductal cell monolayers (in Ham's F10). Additional embryos were cultured exclusively in Ham's F10 or on cat oviductal cell monolayers. Fertilization rates were lower (P < 0.05) on oviductal cells but cell number per embryo was similar (P > 0.05) in all treatments. Blastocyst formation was lower (P < 0.05) on oviductal cells than in mKrb-Ham's F10 treatment and was <20% in all treatments. In summary, while in vitro fertilization-derived cat embryos develop to morulae under a variety of culture conditions, the morula-to-blastocyst developmental block was minimally responsive to alterations in energy substrate and medium complexity or fluctuations in their temporal availability. In addition, oviductal cell culture, alone or in combination with other culture variations, was ineffective in overcoming the developmental block. (C) 1996 Wiley-Liss, Inc.