title: Substrate utilization in porcine embryos cultured in NCSU23 and G1.2/G2.2 sequential culture media.
abstract: Embryo metabolism is an indicator of viability and, therefore, efficiency of the culture medium. Currently, little is known regarding porcine embryo metabolism. The objective of our study was to evaluate glucose and pyruvate uptake and lactate production in porcine embryos cultured in two different media systems. Oocytes were matured and fertilized according to standard protocols. Embryos were allocated randomly into two culture treatments, NCSU23 medium or G1.2/G2.2 sequential culture media 6-8 h post-insemination (hpi). Embryo substrate utilization was measured at the two-cell (24-30 hpi), 8-cell (80 hpi), morula (120 hpi), and blastocyst (144 hpi) stages using ultramicrofluorimetry. Glucose uptake was higher (P < 0.05) in two-cell embryos cultured in G1.2 than in NCSU23 medium (4.54 +/- 0.71, 2.16 +/- 0.87 pmol/embryo/h, respectively). Embryos cultured in G1.2/G2.2 produced significantly more lactate than those in NCSU23 at the eight-cell stage (9.41 +/- 0.71, 4.42 +/- 0.95 pmol/embryo/hr, respectively) as well as the morula stage (11.03 +/- 2.31, 6.29 +/- 0.77 pmol/embryo/hr, respectively). Pyruvate uptake was higher (P < 0.05) in morula cultured in G1.2/G2.2 versus NCSU23 (22.59 +/- 3.92, 11.29 +/- 1.57 pmol/embryo/h, respectively). Lactate production was greater (P < 0.05) in blastocysts cultured in G1.2/G2.2 (38.13 +/- 15.94 pmol/embryo/h) than blastocysts cultured in NCSU23 (8.46 +/- 2.38 pmol/embryo/h). Pyruvate uptake was also greater in blastocysts cultured in G1.2/G2.2 (24.3 +/- 11.04) than those in NCSU23 (11.30 +/- 2.70). When cultured in NCSU23 medium, two- and eight-cell embryos utilized less glucose than morulae and blastocysts, and two-cell embryos produced less lactate than blastocysts (P < 0.05). In G1.2/G2.2 media, two-cells took up less pyruvate than morulae or blastocysts, while blastocysts produced more lactate and utilized more glucose than two-cell, eight-cell and morula stage embryos (P < 0.05). As in other species, glycolysis appears to be the primary metabolic pathway in post-compaction stage porcine embryos. Culture medium composition affects not only substrate uptake, but also metabolic pathways by which these substrates are utilized in porcine embryos at several developmental stages.
authors.affiliation: Department of Animal Sciences, Purdue University, West Lafayette, Indiana 47907-1151, USA.
authors.forename: A P
authors.initials: AP
authors.lastname: Gandhi
authors.forename: M
authors.initials: M
authors.lastname: Lane
authors.forename: D K
authors.initials: DK
authors.lastname: Gardner
authors.forename: R L
authors.initials: RL
authors.lastname: Krisher
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002970:Cleavage Stage, Ovum / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry*; D046508:Culture Techniques; D004622:Embryo, Mammalian / Q000378:metabolism*; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro; D005947:Glucose / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism*; D008297:Male; D009028:Morula / Q000378:metabolism; D019289:Pyruvic Acid / Q000378:metabolism*; D011897:Random Allocation; D013552:Swine / Q000196:embryology*
chemical_list: D003470:Culture Media; D019344:Lactic Acid; D019289:Pyruvic Acid; D005947:Glucose