Impact of adding 5.5 mM glucose to SOF medium on the development, metabolism and quality of in vitro produced bovine embryos from the morula to the blastocyst stage.
Although toxic for early stages of embryo development, glucose is a physiological metabolic substrate at the morula and blastocyst stages. We evaluated the effect of adding 5.5 mM glucose from the morula stage on bovine blastocyst development and quality. In vitro matured and fertilised bovine oocytes were cultured in modified Synthetic Oviduct Fluid medium containing 5% fetal calf serum, but without added glucose, up to day 5 post-insemination (pi). Morulae were selected and further cultured in the presence or absence of 5.5 mM glucose. Blastocyst and hatched blastocyst rates were recorded. Oxygen, glucose and pyruvate uptakes as well as lactate release were evaluated. The quality of the resulting blastocysts was evaluated by the cell allocation to the inner cell mass (ICM) and trophectoderm (TE) and by the apoptotic index. Adding glucose increased the blastocyst rate at day 8 pi (80% vs 65%) but had no impact on hatching rate (25% vs 28%). A 22% decrease in oxygen uptake was observed in the presence of glucose, concomitant with an increase in lactate release, although no change was observed in pyruvate uptake. A slight decrease in blastocyst cell number was observed at day 7 in the presence of glucose while neither the ICM/TE cell ratio nor the apoptotic index were affected. In conclusion, adding 5.5 mM glucose from the morula stage has a limited impact on blastocyst rate and quality although important modifications were observed in embryo metabolism. It remains to be determined whether those modifications could influence embryo viability after transfer.
Donnay
I
I
Veterinary Unit, Catholic University of Louvain, Louvain-la-Neuve, Belgium. donnay@vete.ucl.ac.be
Feugang
J M
JM
Bernard
S
S
Marchandise
J
J
Pampfer
S
S
Moens
A
A
Dessy
F
F
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D003470:Culture Media; D005314:Embryonic and Fetal Development / Q000187:drug effects* / Q000502:physiology; D004734:Energy Metabolism / Q000187:drug effects; D005307:Fertilization in Vitro; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D066298:In Vitro Techniques; D019344:Lactic Acid / Q000378:metabolism; D009028:Morula / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects; D019289:Pyruvic Acid / Q000378:metabolism
D003470:Culture Media; D019344:Lactic Acid; D019289:Pyruvic Acid; D005947:Glucose