title: Supplementation of maturation medium with L-carnitine improves cryo-tolerance of bovine in vitro matured oocytes.
abstract: The objective was to determine the effects of adding L-carnitine (an enhancer of lipid metabolism) during IVM, on cryotolerance and developmental competence of bovine oocytes. Oocytes matured in the absence (control) or presence (0.6 mg/mL) of L-carnitine were subjected to IVF and embryo culture after Cryotop vitrification or nonvitrification at the metaphase stage of the second meiotic cell division. Cleavage and blastocyst formation rates, and inner cell mass and trophectoderm cell numbers were determined. Also, ATP content in IVM oocytes was measured and intracellular lipid droplets were observed (Nile red staining and confocal microscopy). L-carnitine had no significant effect on the rate of matured oocytes. Vitrification reduced (P < 0.05) mean (±SEM) rates of live oocytes both in control (80.6 ± 1.9%) and L-carnitine groups (82.7 ± 5.1%) compared with nonvitrified oocytes (100%). After IVF, cleavage rates of vitrified control and L-carnitine groups (56.5 ± 3.9% and 62.8 ± 5.1%, respectively) were significantly lower than those in nonvitrified control and L-carnitine groups (83.9 ± 4.2% and 84.3 ± 1.3%). After vitrification, blastocyst formation rate in the L-carnitine group (54.4 ± 5.2%) was significantly higher compared with the control (34.9 ± 4.4%), and did not significantly differ from those in nonvitrified control and L-carnitine groups (52.1 ± 4.2% and 52.8 ± 3.0%). The numbers and ratio of inner cell mass and trophectoderm cells in blastocysts did not differ significantly among groups. The ATP content in L-carnitine-treated oocytes tended to be higher compared with the control. Vitrification did not reduce ATP content in oocytes, irrespective of L-carnitine treatment. Treatment with L-carnitine dislocated lipid droplets from the peripheral area to the inner cytoplasm. In conclusion, L-carnitine supplementation during IVM redistributed lipid droplets in oocytes; if they survived vitrification, their developmental competence was similar to that of nonvitrified oocytes.
authors.affiliation: NARO Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan.
authors.forename: Vibuntita
authors.initials: V
authors.lastname: Chankitisakul
authors.forename: Tamas
authors.initials: T
authors.lastname: Somfai
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Inaba
authors.forename: Mongkol
authors.initials: M
authors.lastname: Techakumphu
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagai
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002331:Carnitine / Q000008:administration & dosage*; D002417:Cattle; D002478:Cells, Cultured; D015925:Cryopreservation / Q000662:veterinary; D003451:Cryoprotective Agents / Q000008:administration & dosage*; D003470:Culture Media; D046149:Embryo Culture Techniques / Q000662:veterinary; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D009865:Oocytes / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*
chemical_list: D003451:Cryoprotective Agents; D003470:Culture Media; D000255:Adenosine Triphosphate; D002331:Carnitine