Consequences of exposure to serum, with or without vitamin E supplementation, in terms of the fatty acid content and viability of bovine blastocysts produced in vitro.
To determine whether serum supplementation influenced fatty acid content of bovine blastocysts and whether vitamin E addition to culture medium containing serum could improve development in vitro, cleaved eggs were cultured in synthetic oviduct fluid supplemented with bovine serum albumin (BSA, 0.4% w/v, fraction V) (SVBSA), fetal calf serum (FCS, 10% v/v) (SFCS) or FCS (10% v/v) plus 100 micro M vitamin E (SFCS + E). Blastocyst yields were recorded and fatty acid composition was determined by gas chromatography. Day 7 blastocysts were incubated with [2-(14)C] pyruvate for 3 h and then fixed for cell counts. Yields of good quality blastocysts were greatest from cleaved eggs cultured in serum-free conditions (P < 0.01). In the presence of serum, supplementation with vitamin E increased both total and good quality blastocyst yields (P < 0.01). Presence of serum increased fatty acid content (mean +/- SEM) of blastocysts (SVBSA v. SFCS = 57 +/- 2 v. 74 +/- 2 ng embryo(-1); P < 0.001). In contrast, pyruvate metabolism was greater in blastocysts produced without serum (27 +/- 3 v. 21 +/- 3 picomoles embryo(-1) 3h(-1); P < 0.01) but, on a per cell basis, no differences were detected. Addition of vitamin E to the serum-supplemented formulation did not alter either the fatty acid content (73 +/- 2 ng embryo(-1)) or pyruvate metabolism index (19 +/- 1 pmol embryo(-1) 3h(-1)) of SFCS + E blastocysts. Thus, despite lipid accumulation, supplementary vitamin E improved blastocyst yields in embryos exposed to serum.
Reis
A
A
Scottish Agricultural College, Animal Biology Division, Craibstone Estate, Bucksburn, Aberdeen, UK.
Rooke
J A
JA
McCallum
G J
GJ
Staines
M E
ME
Ewen
M
M
Lomax
M A
MA
McEvoy
T G
TG
D000818:Animals; D001755:Blastocyst / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D002417:Cattle; D018929:Cell Culture Techniques; D002470:Cell Survival; D002970:Cleavage Stage, Ovum / Q000187:drug effects / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D005227:Fatty Acids / Q000032:analysis* / Q000378:metabolism; D005260:Female; D019289:Pyruvic Acid / Q000032:analysis / Q000378:metabolism; D044967:Serum / Q000737:chemistry; D014810:Vitamin E / Q000032:analysis / Q000494:pharmacology*
D003470:Culture Media; D005227:Fatty Acids; D014810:Vitamin E; D019289:Pyruvic Acid