大鼠卵巢颗粒细胞的分离方法

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.