Wolski Katja M, Feig Caroline, Kirchhoff Christiane, Cameron Don F
Department of Pathology and Cell Biology, University of South Florida, College of Medicine, Tampa, FL 33612, USA.
Asian J Androl. 2007 May;9(3):312-20. doi: 10.1111/j.1745-7262.2007.00256.x.
To determine the effectiveness of the sk11, sk9 and sk11 TNUA5 Sertoli cell lines in binding germ cells in vitro.
The immortalized Sertoli cell lines sk9, sk11 and sk11 TNUA5 were used in co-culture experiments with germ cells in media with or without reproductive hormones and incubated for 44 h at 32 degrees . The number of germ cells bound to Sertoli cells was then determined and statistically analyzed. Western blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR) studies were employed to investigate the presence of cell adhesion proteins and follicle stimulating hormone (FSH) receptor, respectively.
No statistical difference between the number of bound step-8 spermatids and bound pre-step 8 spermatids on Sertoli cells from any of the cell lines existed. After the addition of germ cells, Sertoli cells showed more lipid accumulation in their cytoplasm, indicating active phagocytosis. Western blot analysis in the sk11 TNUA5 line indicated the expression of N-cadherin. FSH-only and testosterone-only treatments increased N-cadherin expression, regardless of germ cell addition. The addition of germ cells to the sk11 TNUA5 Sertoli cells increased the expression of espin, as did the addition of FSH with germ cells. RT-PCR studies of the sk11 TNUA5 cells indicated that the mRNA for FSH receptor decreased with successive passages.
In vitro binding between isolated germ cells and sk9, sk11 or sk11 TNUA5 Sertoli cells is not feasible, and therefore these cell lines are not useful for the in vitro investigation of Sertoli-germ cell interactions and primary Sertoli cell isolates must still be used.
确定sk11、sk9和sk11 TNUA5这三种台湾艺术大学（TNUA）的支持细胞系在体外结合生殖细胞的有效性。
将永生化的支持细胞系sk9、sk11和sk11 TNUA5与生殖细胞在含有或不含有生殖激素的培养基中进行共培养实验，并在32摄氏度下孵育44小时。然后确定与支持细胞结合的生殖细胞数量并进行统计分析。采用蛋白质免疫印迹分析和逆转录聚合酶链反应（RT-PCR）研究分别检测细胞粘附蛋白和促卵泡激素（FSH）受体的存在情况。
任何细胞系的支持细胞上结合的8期精子细胞数量与结合的8期前精子细胞数量之间均无统计学差异。加入生殖细胞后，支持细胞的细胞质中出现更多脂质积累，表明存在活跃的吞噬作用。sk11 TNUA5细胞系的蛋白质免疫印迹分析表明N-钙黏蛋白表达。无论是否添加生殖细胞，单独使用FSH和单独使用睾酮处理均增加了N-钙黏蛋白的表达。向sk11 TNUA5支持细胞中添加生殖细胞增加了espin的表达，添加FSH与生殖细胞时也有同样效果。对sk11 TNUA5细胞的RT-PCR研究表明，随着传代次数增加，FSH受体的mRNA减少。
分离的生殖细胞与sk9、sk11或sk11 TNUA5支持细胞之间的体外结合不可行，因此这些细胞系对于支持细胞-生殖细胞相互作用的体外研究无用，仍必须使用原代分离的支持细胞。
