Lee Hak-Mo, Oh Byoung Chol, Lim Dong-Pyo, Lee Dong-Sup, Cho Jaejin, Lee Gene, Lee Jeong Ryul
Department of Thoracic and Cardiovascular Surgery, Seoul National University, Seoul, Korea.
Xenotransplantation. 2007 Mar;14(2):112-8. doi: 10.1111/j.1399-3089.2007.00366.x.
An understanding of the main mechanism that determines the ability of immune privilege related to Sertoli cells (SC) will provide clues for promoting a local tolerogenic environment. In this report, we established neonatal porcine SC line and evaluated their characteristics.
SC line was established following the transfection of primary SC (NPSC) from the testis of neonatal pig with plasmid pRNS-1 carrying genes for neomycin resistance and the SV40 large T antigen. Immunohistochemistry and RT-PCR were performed to evaluate the character of immortalized SC lines.
Our immortalized SC line (iPS) proliferated stably and had a phenotype similar to NPSC, as indicated by the immunoexpression of follicle stimulating hormone receptor (FSHR), and mRNA expression of androgen receptor (AR), and Wilms' tumor antigen (WT1). Interestingly, NPSC and iPS expressed mRNA of complement regulatory proteins (CRP) such as membrane cofactor protein (CD46), decay accelerating factor (DAF or CD55), and protectin (CD59), but CD59 mRNA expression was negligible in iPS.
These results suggest that iPS, immortalized by the introduction of SV40 T, retain their original characteristics, except for the relatively low expression of CD59, and that they may be useful for future in vitro and in vivo studies of immune privilege mechanisms related to SC.
了解决定支持细胞（SC）相关免疫豁免能力的主要机制，将为促进局部耐受环境提供线索。在本报告中，我们建立了新生猪支持细胞系并评估了它们的特性。
用携带新霉素抗性基因和SV40大T抗原基因的质粒pRNS-1转染新生仔猪睾丸的原代支持细胞（NPSC），建立支持细胞系。采用免疫组织化学和逆转录-聚合酶链反应（RT-PCR）评估永生化支持细胞系的特性。
我们的永生化支持细胞系（iPS）稳定增殖，具有与NPSC相似的表型，卵泡刺激素受体（FSHR）的免疫表达、雄激素受体（AR）和肾母细胞瘤抗原（WT1）的mRNA表达均表明了这一点。有趣的是，NPSC和iPS表达补体调节蛋白（CRP）如膜辅因子蛋白（CD46）、衰变加速因子（DAF或CD55）和保护素（CD59）的mRNA，但iPS中CD59 mRNA表达可忽略不计。
这些结果表明，通过引入SV40 T永生化的iPS保留了其原始特性，但CD59表达相对较低，并且它们可能有助于未来对与支持细胞相关的免疫豁免机制进行体外和体内研究。
