权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

次世代型人工臓器設計のための腹膜機能の測定とモデル化

下一代人工器官设计的腹膜功能测量和建模

基本信息

批准号：
07044169
负责人：
HORIUCHI Takashi
金额：
$ 6.4万
依托单位：
University of East Asia
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

The hybrid-type peritoneal membrane model is a promising tool to understand not only solute and water transport via the peritoneum but also fibrotic process due to endogenous and/or exogenous stimuli. There exist, however, technological requirements to be overcome such as a cloning of peritoneal fibroblasts and their 3D cultures in the extra cellular matrices.In this study we have surveyed the 3 dimensional culture of the peritoneal resident cells to reconstruct the peritoneum in vitro and found few research activitie on it except for some studies of host defense mechanizms using cultured peritoneal cells. Modifying their techniques we established a time elapsed-differential subculture (t-DSC), regulating contact time to the culture plastics, to separate fibroblasts from sub-confluent primary cultured peritoneal cells. Establishing PRFB (1*10^5/ml) in collagen gels composed of increasing concentrations of collagen resulted in an inverse correlation with gel contraction while increasing the PRFB cell density within the collagen gels (2mg/ml) resulted in a direct correlation with collagen gel contraction. Addition of the transcriptional inhibitor actinomycin D to 3D-RPFB cultures resulted in a dose dependent inhibition of gel contraction. Gel contraction, however, was not inhibited following the addition of the translation inhibitor, puromycin and similarly no significant effect was seen following incubation with 2,2'-bipyridyl. From these results, it could be hypothesized that mechanisma of collagen gel contraction may relate to change in solute permeability of the peritoneal patients in a long-term peritoneal dialysis. We found an expression of TGF-beta mRNA expresssion in this experimental setting which could indicate an expression of integrin.It is concluded that in vittro 3D peritoneal model is promissing method to understand mechanism of not only solute permeability through the peritoneum but also fibrotic process.

混合型腹膜模型是一个很有前途的工具，了解不仅溶质和水通过腹膜的运输，但也纤维化过程中由于内源性和/或外源性刺激。然而，目前还存在一些技术上的问题，如腹膜成纤维细胞的克隆及其在细胞外基质中的三维培养等，本研究对腹膜驻留细胞的三维培养进行了综述，发现除了利用培养的腹膜细胞进行宿主防御机制的研究外，对腹膜驻留细胞的三维培养的研究很少。修改他们的技术，我们建立了一个时间流逝的差异继代培养（t-DSC），调节接触时间的培养塑料，分离成纤维细胞亚融合的原代培养的腹膜细胞。在由增加浓度的胶原蛋白组成的胶原蛋白凝胶中建立PRFB（1*10^5/ml）导致与凝胶收缩的负相关，而增加胶原蛋白凝胶内的PRFB细胞密度（2 mg/ml）导致与胶原蛋白凝胶收缩的直接相关。在3D-RPFB培养物中加入转录抑制剂放线菌素D导致凝胶收缩的剂量依赖性抑制。然而，在加入翻译抑制剂嘌呤霉素后，凝胶收缩没有受到抑制，并且类似地，在与2，2 '-联吡啶孵育后没有观察到显著效果。根据这些结果，可以假设胶原凝胶收缩的机制可能与腹膜透析患者在长期腹膜透析中溶质渗透性的变化有关。我们发现TGF-β mRNA的表达在这个实验环境中，这可能表明integrin的表达。结论是，在体外3D腹膜模型是有前途的方法来理解不仅溶质渗透通过腹膜的机制，而且纤维化过程。