COCs exhibited different diffusion situation between control and treatment group: the diffusion of 200 µM LIMKi 3 treated-COCs was significantly weaker than the control group. In the standard of COCs surrounded by the cumulus cell layers, we counted the proportion of diffusion and found that the rate of good diffusion was decreased significantly (40.05 ± 3.03% vs. 20.50 ± 4.05%; p < 0.05). Oocytes were then obtained from COCs after hyaluronidase treatment, we found that few oocytes extruded the polar body after 200 µM LIMKi 3 treatment.
|Preparation method||For LIMK1/2 inhibitor treatment, stock LIMKi 3 (50 mM in dimethylsulfoxide (DMSO)) was diluted in M199 to final concentrations of 50, 100, 150 and 200 µM. A control group was cultured in DMSO at the same relative concentration of solvent. COCs were cultured with LIMKi 3 to evaluate its effects on oocyte maturation. COCs were denuded of their cumulus cells by gentle pipetting with 0.1% (w/v) hyaluronidase.|
|Incubation time||44 h|
|Animal models||NF1+/− mice|
|Formulation||DMEM × 1 containing 5% FCS|
|Dosages||0, 25 or 50 μM|
|Body Surface Area (m2)||0.007||0.025||0.15||0.05||0.02||0.5|
|Animal A (mg/kg) = Animal B (mg/kg) multiplied by||Animal B Km|
|Animal A Km|
For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.
|Solubility||34 mg/mL in DMSO|
LIMK1/2 inhibitor LIMKi 3 suppresses porcine oocyte maturation.
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