Isolation method of Leydig cells from mature male Djungarian hamsters (Phodopus sungorus) and their steroidogenic activity in vitro
Isolation method of Leydig cells from mature male Djungarian hamsters (Phodopus sungorus) and their steroidogenic activity in vitro
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成熟雄性准加仓鼠 (Phodopus sungorus) 间质细胞的分离方法及其体外类固醇生成活性
DOI：
10.1111/j.1439-0272.1999.tb01403.x
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发表时间：
1999
期刊：
影响因子：
2.4
通讯作者：
A. Lerchl
中科院分区：
文献类型：
--
作者：
M. Niedziela;A. Lerchl
对于解决Leydig细胞功能的研究，分离的细胞通常比完整的动物更适合。在这里，Leydig细胞的分离程序从成年雄性Djungarian仓鼠（Photopus sungorus）的描述。使用涉及酶促解离和Percoll梯度离心的程序分离细胞。对于每个实验，从6只动物中获得约4.4 × 106个Leydig细胞。细胞表现出较高的类固醇反应生理（羊促黄体激素（oLH）和人绒毛膜促性腺激素（hCG））和非生理（毛喉素）刺激体外。通过台盼蓝拒染法评估，约98%的细胞存活，通过3β-羟基类固醇脱氢酶活性检测，纯度为80 - 95%。在相差显微镜下还通过亮黄色晕圈识别了间质细胞。它们含有大量脂滴，核圆，核仁突出。细胞对oLH、hCG和毛喉素的反应是以剂量依赖性方式增加睾酮的产生。这些研究中的剂量反应曲线表明，Djungarian仓鼠的Leydig细胞发生脱敏，可能是由于其LH/CG受体的下调。
For studies addressing the functions of Leydig cells, isolated cells are often better suited than intact animals. Here, the isolation procedure of Leydig cells from adult male Djungarian hamsters (Phodopus sungorus) is described. Cells were isolated using a procedure involving enzymatic dissociation and Percoll‐gradient centrifugation. For each experiment, ≅4.4 × 106 Leydig cells from six animals were obtained. The cells showed high steroidogenic responsiveness to physiological (ovine luteinizing hormone (oLH) and human chorionic gonadotropin (hCG)) and nonphysiological (forskolin) stimuli in vitro. Approximately 98% of cells were viable as assessed by trypan blue exclusion, and the purity varied from 80 to 95% as tested by 3β‐hydroxysteroid dehydrogenase activity. Leydig cells were also identified by a bright yellow halo under phase‐contrast microscopy. They contained numerous lipid droplets and showed round nuclei and prominent nucleoli. The cells responded to oLH, hCG and forskolin with an increased testosterone production in a dose‐dependent manner. Dose‐response curves in these studies suggest that Leydig cells of Djungarian hamsters undergo desensitization, probably due to down regulation of their LH/CG receptors.
影响因子：
3.6
作者：
KLINEFELTER, GR;HALL, PF;EWING, LL
通讯作者：
EWING, LL