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1. Reprod Fertil Dev. 2011 Dec;24(1):194-5.

165 the effects of L-carnitine and linoleic Acid albumin supplementation on the
development and cryosurvival of bovine in vitro-matured/in vitro-fertilized
embryos in in vitro culture medium.

Miyashita S, Inaba Y, Somfai T, Geshi M, Nagai T, Koyama H, Dochi O.

Department of Dairy Science, Rakuno Gakuen University, Ebetsu, Hokkaido, Japan;

The objective of this study was to investigate the effects of the supplementation
of a lipid metabolism inducer, L-carnitine (LC) and a membrane stabilizer,
linoleic acid albumin (LAA), on the developmental competence and cryosurvival of
bovine in vitro-matured/in vitro-fertilized embryos in in vitro culture medium.
Cumulus-oocyte complexes collected from the ovaries of slaughtered cattle were
matured for 20h in TCM-199 supplemented with 5% calf serum (CS) and 0.02AUmL(-1)
of FSH at 38.5°C in an atmosphere of 5% CO(2) in air. After IVF (Day 0),
presumptive zygotes were cultured in CRlaa containing 5% CS at 38.5°C in an
atmosphere of 5% CO(2), 5% O(2) and 90% N(2) for 9 days. The culture medium was
supplemented with 0.6mgmL(-1) of LC (LC group; n=180) or with 0.25mgmL(-1) of LAA
(LAA group; n=180) or with both LC and LAA (LC+LAA group; n=180) or without LC
and LAA (control; n=178). The cleavage rates were recorded on Day 2 and the
blastocyst formation rates were recorded on Day 7 to 9. Expanded blastocysts
harvested on Day 7 and 8 (LAA group: n=31; LC group: n=29; LC+LAA group: n=25;
control group: n=33) were used for freezing in modified PBS supplemented with
1.5M ethylene glycol, 0.1M sucrose and 20% CS. After thawing, they were cultured
in TCM-199 supplemented with 20% FBS and 0.1mM β-mercaptoethanol at 38.5°C under
5% CO(2) in air for 72h. The rates of re-expansion, hatching and formation of
hatched blastocysts were determined at 24, 48 and 72h after thawing,
respectively. The rates of cleavage and blastocyst formation were expressed as
mean±s.e.m. and analysed by ANOVA. The post-thaw survival rates of frozen embryos
were analysed by chi-square test. The cleavage rate in the control group
(69.1±2.5%) was significantly lower than that in the LAA (81.8±3.8%) and LC+LAA
groups (77.9±1.4%) but did not differ from that in the LC group (73.8±2.4%). The
blastocyst formation rate in the control group (21.7±2.8%) was significantly
lower (P<0.05) than those in the LAA and LC+LAA groups (33.5±2.8% and 31.4±2.4%,
respectively), but it did not differ significantly from that of the LC group
(32.1±3.3%) despite a strong tendency (P=0.06). There were no significant
differences among the control, LC, LAA and the LC+LAA groups in post-thaw
re-expansion rates (66.7, 75.9, 67.7 and 76.0%, respectively), hatching rates
(48.5, 69.0, 58.1 and 64.0%, respectively) and rates of formation of hatched
blastocysts (51.5, 62.1, 61.3 and 64.0%, respectively). These results indicate
that the addition of LC and LAA to the medium for in vitro culture of in
vitro-matured/in vitro-fertilized bovine embryos improved their ability to
develop to the blastocyst stage; however, the effects on the freezing tolerance
were not verified.

PMID: 22394888 [PubMed - in process]