The effects of dimethyl-sulfoxide on the in vitro maturation and fertilization of bovine oocytes and the subsequent development

The Effect of Various Concentrations of Taurine during In vitro Fertilization on the Development of Bovine Embryos Fertilized with Spermatozoa from Three Different Bulls

“…During IVF, 0, 1, 10, 50 and 100 mM taurine (No.T-8691, Sigma-Aldrich) were added to the IVF medium (the culture volume of the IVF medium was adjusted to 5 μl/COC in all groups). After 6 hrs' incubation, the COCs were washed with TCM-199 and transferred to CR1aa (Rosenkrans and First, 1991) supplemented with 50 μM DMSO (Tsuzuki et al, 1998), 1% CS and antibiotics, as mentioned above. For the other IVF method, we used 100 μg/ml heparin sodium (H) only for 24±2 h (Tsuzuki et al, 2005) to induce sperm capacitation with modification from previous papers (Fukui et al, 1990;Gordon, 1994;Saeki et al, 1995;Guyader-Joly et al, 1998;Chohan and Hunter, 2004) because in our preliminary study, when the oocytes were fertilized with the same bull's semen in the presence of 1, 10 and 100 μg/ml H for the same period (24±2 h), the development rates up to the blastocyst stage in the 100 μg/ml H group in all bulls were higher than those of the other H concentration groups.…”

“…The cultural volume in both the CH and H groups was adjusted to 5 μl/COC during IVF. On 3 day from the onset of IVF, the culture medium was changed to CRlaa supplemented with 50 μM DMSO (Tsuzuki et al, 1998), 15% CS and antibiotics with 5 μl/COC of the culture medium, and the cleavage rates from 2 to 16-cells stage were observed by an inverted phase contrast microscope. At 7-8 days from the onset of IVF, the blastocyst formation rates in each group were also observed.…”

Effects of Cytochalasin-D on the Maintenance of Blastocoels of Bovine Blastocysts Produced In Vitro

“…F-2293, Sigma, MO, USA), 50 µM dimethyl-sulfoxide (DMSO, No. 134-07, Nacalai Tesque, Kyoto, Japan, [11]), 5% (V/V) calf serum (CS, Gibco BRL Products), a n d a n t i b i o t i c s ( 9 8 U / m l p e n i c i l l i n , 9 8 µ g / m l streptomycin, Katayama Chemical Co. and 100 µg/ml dibekacin sulfate, Meiji Seika Co., Tokyo, Japan) for 23-24 hr under 5% CO 2 , 95% air and 100% humidity.…”