Three-week old female mice were injected with 5 I.U. PMSG to induce superovulation. Forty-four hours after injection, the mice were euthanized and the ovaries dissected into media containing HEPES and 1 μM cilostamide (Millipore, 231085) (HC media). Antral follicles were punctured, allowing release of cumulus-oocyte complexes (COCs). Repeated aspiration through a glass pipette allowed for removal of the surrounding cumulus cells. Denuded oocytes were maintained at prophase I arrest (Pro I) in MEM Alpha (Gibco, 12561-056) supplemented with sodium pyruvate (Gibco, 11360070) and penicillin-streptomycin (Gibco, 15140122) in addition to 1 μM cilostamide (αC media) at 37°C under 5% CO2. If indicated, oocytes were transferred to cilostamide-free MEM Alpha (α media), allowed to mature, and collected at various time points. Where specified, oocytes were treated with 5 μM dinaciclib (Selleckchem, SCH727965), 10 μM Ro-3306 (Selleckchem, S7747), or 10 mM Rp-cAMPS.
Cilostamide
Manufactured by Thermo Fisher Scientific
Cilostamide is a laboratory reagent used in research applications. It is a cyclic nucleotide phosphodiesterase inhibitor. The core function of Cilostamide is to inhibit the enzymatic activity of phosphodiesterases, which are responsible for the breakdown of cyclic nucleotides such as cAMP and cGMP within cells.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Dinaciclib, by Selleck Chemicals (1 mentions)
Alpha mem, by Thermo Fisher Scientific (1 mentions)
Ro 3306, by Selleck Chemicals (1 mentions)
Noradrenaline, by Merck Group (1 mentions)
Milrinone, by Merck Group (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Agarose, by Thermo Fisher Scientific (1 mentions)
Fibrinogen, by Merck Group (1 mentions)
Thrombin, by Thermo Fisher Scientific (1 mentions)
Isoprenaline, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Harvard Bioscience (1 mentions)
Insulin, by Merck Group (1 mentions)
Pluronic f 127, by Merck Group (1 mentions)
Aprotinin, by Merck Group (1 mentions)
Collagenase 2, by Worthington (1 mentions)
Milrinone, by Thermo Fisher Scientific (1 mentions)
3 protocols using cilostamide
1
Oocyte Maturation and Arrest Modulation
2
Cardiac Tissue Engineering Protocols
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The suppliers of the materials used in these experiments are as follows: Sigma‐Aldrich supplied isoprenaline (I6504); noradrenaline (A7257); milrinone (M4659); tadalfil (Y0001417); Pluronic F127, (P2443); fibrinogen, (F8630); insulin, (I9278); aprotinin, (A1153). R&D Systems supplied cilostamide (091510) and rolipram (90510) and Gibco supplied penicillin/streptomycin (15140) and Geltrex (A1413302). Other sources of compounds were: EDTA: Roth (8043.2); collagenase II, Worthington (LS004176); anti‐cardiac troponin T‐FITC, recombinant human IgG1, clone REA400, 1:10, Miltenyi Biotec (130–106‐687); agarose, Invitrogen (15510–027); thrombin, Peprotech (100–21); horse serum, Life technologies (26050088); DMEM, Biochrom (F0415).
3
Preparing Mouse Cumulus-Oocyte Complexes
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Female mice were injected with 5 I.U. pregnant mare serum gonadotropin (PMSG; MyBioSource) to induce follicle growth and sacrificed 44 h after injection. The ovaries were dissected, and COCs were collected in media containing HEPES-modified minimum essential medium Eagle (Sigma-Aldrich, M2645) supplemented with 0.23 mM sodium pyruvate (Gibco, 11360070), penicillin–streptomycin (Genesee Scientific Corporation (GenClone), 25-512), and 3 mg/ml BSA (Sigma-Aldrich, SIAL-A3311), 6 mM sodium bicarbonate (JT-Baker, 3506-1), and a PDE inhibitor (1 µM cilostamide (Calbiochem, 231085) or 2 μM milrinone (Enzo ALK-270-083)). Aspiration through a glass pipette allowed for the removal of the surrounding cumulus cells. Denuded oocytes were maintained at the GV stage in α-MEM (Gibco, 12561-056) supplemented with 0.23 mM sodium pyruvate, penicillin–streptomycin, and 1 µM cilostamide (or 2 μM milrinone) at 37 °C under 5% CO2. For the collection of in vivo MII oocytes, female mice were injected with 5 I.U. of PMSG followed by 10 I.U. human chorionic gonadotropin (hCG; Goldline Labs) 48 h after PMSG injection, and after 13 h, sacrificed for ovulated oocytes retrieval from the fallopian tubes.
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