Zhang Xuan, Jiang Xue-wu, Li Jian-hong, Ma Lian, Huang Tian-hua
Department of Pediatric Surgery, The Second Hospital, China.
Zhonghua Nan Ke Xue. 2009 Oct;15(10):872-5.
To establish a primary culture of the testis gubernacular cells of Kunming mice, observe the morphological characteristics of the cells, and explore the effects of exogenous estrogens (EEs) on the development of the testis gubernacula in vitro.
We removed the gubernacula from 3-day-old mice with the surgical magnifier and cultured the gubernacular cells. Then we detected the cell viability by trypan blue and cell morphology by HE staining. The subcultured cells were randomly divided into a blank control, a DMSO (0.1%, v/v) control, and 4 experimental groups (given 0.01, 0.10, 1.00 and 10.00 micdrog/ml of diethylstilbestrol [DES] dissolved in DMSO, respectively). After treated for 12, 24 and 48 hours, the gubernacular cells were observed for morphological changes and proliferation inhibition by CCK-8.
Most of the cultured gubernacular cells were fibroblasts, and a few were epithelioids. The primary cells showed a viability of 85%-90%. Dose- and time-dependent inhibition of cell proliferation was found in the four experimental groups at three different times, with statistically significant differences (P < 0.01).
Gubernacular cells can be cultured in vitro. EEs inhibit the proliferation of gubernacular cells in a dose- and time-dependent manner. An in- sight into the effects EES on cultured gubernacular cells is an effective approach to the study of their influence on the development of the reproductive system.
建立昆明小鼠睾丸引带细胞原代培养体系，观察细胞形态特征，探讨外源性雌激素（EEs）对体外睾丸引带发育的影响。
在手术放大镜下取3日龄小鼠的引带并培养引带细胞。然后用台盼蓝检测细胞活力，用苏木精-伊红（HE）染色观察细胞形态。将传代培养的细胞随机分为空白对照组、二甲基亚砜（DMSO，0.1%，v/v）对照组和4个实验组（分别给予溶于DMSO的0.01、0.10、1.00和10.00μg/ml己烯雌酚[DES]）。处理12、24和48小时后，观察引带细胞的形态变化，并用CCK-8检测增殖抑制情况。
培养的引带细胞大多为成纤维细胞，少数为上皮样细胞。原代细胞活力为85%-90%。在三个不同时间点，四个实验组均发现细胞增殖受到剂量和时间依赖性抑制，差异有统计学意义（P<0.01）。
引带细胞可在体外培养。EEs以剂量和时间依赖性方式抑制引带细胞增殖。深入了解EEs对培养的引带细胞的影响是研究其对生殖系统发育影响的有效途径。
