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        大鼠卵巢颗粒细胞的分离方法

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        大鼠颗粒细胞的分离方法一:

        分离23天的SD大鼠,皮下注射DES(2.5 mg/只),连续三天。于最后一次注射24 h后,颈椎脱臼处死动物。无菌收集两侧的卵巢,置于盛有PBS的EP管中清洗。将PBS清洗的卵巢放在盛有培养基的平皿中,在体视显微镜下,用25号针头刺破卵泡,释放出颗粒细胞。后吸取平皿中的细胞悬液,镜下计数,接种细胞。培养基(McCoy's 5A Medium),加谷氨酰氨,用碳酸氢钠调节PH值为7.2。

        参考文献:
        1.Down-Regulation of Steroidogenic Response to Gonadotropins in Human and Rat Preovulatory Granulosa Cells Involves Mitogen-Activated Protein Kinase Activation and Modulation of DAX-1 and Steroidogenic Factor-1.J Clin Endocrinol Metab May 2003,88:2288-2299.
        2.Activin-A, but not inhibin, regulates 17beta-hydroxysteroid dehydrogenase type 1 activity and expression in cultured rat granulosa cells.J Steroid Biochem Mol Biol, Jul 2000,73(5):203-10.
        3.Synergistic effects of activin and FSH on hyperphosphorylation of Rb and G1/S transition in rat primary granulosa cells.Mol Cell Endocrinol.Nov 2003,210(1-2):31-38.
        4.Induction of Cyclin D2 in Rat Granulosa Cells Requires FSH-dependent Relief from FOXO1 Repression Coupled with Positive Signals from Smad.J Biol Chem, Mar 2005,280:9135-9148.

        大鼠颗粒细胞的分离方法二:

        23天的SD大鼠,皮下注射PMSG10IU 48 h,hCG 10 IU。方法同前。

        参考文献:
        1.Proliferation of Rat Granulosa Cells during the Periovulatory Interval Endocrinology, Jan 2005,146:414-422.
        2.Transient expression of progesterone receptor messenger RNA in ovarian granulosa cells after the preovulatory luteinizing hormone surge OK Park and KE Mayo Mol.Endocrinol.Jul 1991,5:967.
        3.A method for transplantation of luteinizing granulosa cells: evidence for progesterone secretion.Biol Reprod, Dec 1982,27:1261.
        4.Estrogen Receptor-ß Is Critical to Granulosa Cell Differentiation and the Ovulatory Response to Gonadotropins.Endocrinology, Aug 2005,146:3247-3262.
        5.Regulation of Prostaglandin-Endoperoxide Synthase 2 Messenger Ribonucleic Acid Expression in Mouse Granulosa Cells during Ovulation.Endocrinology, Jul 2001,142:3187.

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