Collagen vitrigel membrane useful for paracrine assays in vitro and drug delivery systems in vivo |
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Authors: | Takezawa Toshiaki Takeuchi Tomoyo Nitani Aya Takayama Yoshiharu Kino-Oka Masahiro Taya Masahito Enosawa Shin |
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Institution: | Laboratory of Animal Cell Biology (currently, Transgenic Animal Research Center), National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan. t.takezawa@affrc.go.jp |
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Abstract: | We previously succeeded in converting a soft and turbid disk of type-I collagen gel into a strong and transparent vitrigel membrane utilizing a concept for the vitrification of heat-denatured proteins and have demonstrated its protein-permeability and advantage as a scaffold for reconstructing crosstalk models between two different cell types. In this study, we observed the nano-structure of the type-I collagen vitrigel membrane and verified its utility for paracrine assays in vitro and drug delivery systems in vivo. Scanning electron microscopic observation revealed that the vitrigel membrane was a dense network architecture of typical type-I collagen fibrils. In the crosstalk model between PC-12 pheochromocytoma cells and L929 fibroblasts, nerve growth factor (NGF) secreted from L929 cells passed through the collagen vitrigel membrane and induced the neurite outgrowth of PC-12 cells by its paracrine effect. Also, the collagen vitrigel membrane containing vascular endothelial growth factor (VEGF) showed sustained-release of VEGF in vitro and its subcutaneous transplantation into a rat resulted in remarkable angiogenesis. These data suggest that the collagen vitrigel membrane is useful for paracrine assays in vitro and drug delivery systems in vivo. |
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