コラーゲンビトリゲル膜を利用した組織特異的なヒスタミン応答性血管内皮モデルの開発

抄録

The tightness of the endothelial barrier is tissue-dependent. We expect that tissue-specific barrier function can be induced in immature vascular endothelial cells by co-culturing with tissue-specific cells. The purpose of this study is to construct tissue-specific vascular endothelial models on a collagen vitrigel membrane (CVM) composed of high-density collagen fibrils equivalent to connective tissues in vivo and compare the responsivity to histamine. We used human microvascular endothelial cells (HMVECs) derived from a newborn foreskin. HMVECs were cultured in a CVM chamber with or without human dermal fibroblasts (HDFs), C6 cells (a rat glioma cell line) or HepG2-NIAS cells (a human hepatocellular carcinoma cell line) cultured on the reverse-side of CVM for up to 6 days. The endothelial barrier function was evaluated by transendothelial electric resistance (TEER). TEER values of a HMVEC monolayer were 15-20 Ω/cm² during culture periods. It significantly increased up to 40-60 Ω/cm² by co-culturing with HDFs, C6 cells and HepG2-NIAS cells. Also, TEER value was clearly decreased in the co-culture model composed of HMVECs and HDFs treated with 1 μM histamine while HMVEC monolayer model showed slight response to 100 μM histamine. Now we are investigating tissue-specific responses to histamine among the models.