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

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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.