Open AccessA new perfusion culture method with a self-organized capillary network
Author(s) -
Kei Sugihara,
Yoshimi Yamaguchi,
Shiori Usui,
Yuji Nashimoto,
Sanshiro Hanada,
Etsuko Kiyokawa,
Akiyoshi Uemura,
Ryuji Yokokawa,
Koichi Nishiyama,
Takashi Miura
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0240552
Subject(s) - perfusion , extravasation , microbiology and biotechnology , cell culture , capillary action , angiogenesis , vascular network , process (computing) , biology , chemistry , biomedical engineering , pathology , anatomy , computer science , medicine , immunology , materials science , cancer research , genetics , composite material , operating system
A lack of perfusion has been one of the most significant obstacles for three-dimensional culture systems of organoids and embryonic tissues. Here, we developed a simple and reliable method to implement a perfusable capillary network in vitro. The method employed the self-organization of endothelial cells to generate a capillary network and a static pressure difference for culture medium circulation, which can be easily introduced to standard biological laboratories and enables long-term cultivation of vascular structures. Using this culture system, we perfused the lumen of the self-organized capillary network and observed a flow-induced vascular remodeling process, cell shape changes, and collective cell migration. We also observed an increase in cell proliferation around the self-organized vasculature induced by flow, indicating functional perfusion of the culture medium. We also reconstructed extravasation of tumor and inflammatory cells, and circulation inside spheroids including endothelial cells and human lung fibroblasts. In conclusion, this system is a promising tool to elucidate the mechanisms of various biological processes related to vascular flow.