Cellular control of tissue architectures using a three-dimensional tissue fabrication technique.
Tissue engineering seeks to provide regenerated tissue architectures in vitro but has not yet successfully created thick, highly vascularized, multi-functional tissues replicating native structure. We describe a novel method to fabricate pre-vascularized tissue equivalents using multi-layered cultures combining micro-patterned endothelial cells as vascular pre-cursors with fibroblast monolayer sheets as tissue matrix. Stratified tissue equivalents are constructed by alternately layering fibroblast monolayer sheets with patterned endothelial cell sheets harvested from newly developed thermo-responsive micro-patterned surfaces alternating 20 microm-wide cell-adhesive lanes with 60 microm non-adhesive zones. Cell culture substrates covalently grafted with different thermo-responsive polymers permit spatial switching of cell adhesion and detachment using applied small temperature changes. Endothelial cell patterning fidelity was maintained within the multi-layer tissue constructs after assembly, leading to self-organization into microvascular-like networks after 5-day tissue culture. This novel technique holds promise for the study of cell-cell communications and angiogenesis in reconstructed, three-dimensional environments as well as for the fabrication of tissues with complex, multicellular architecture.
Tsuda
Yukiko
Y
Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, Japan.
Shimizu
Tatsuya
T
Yamato
Masayuki
M
Kikuchi
Akihiko
A
Sasagawa
Tadashi
T
Sekiya
Sachiko
S
Kobayashi
Jun
J
Chen
Guoping
G
Okano
Teruo
T
D001672:Biocompatible Materials / Q000737:chemistry; D002448:Cell Adhesion; D002478:Cells, Cultured; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D007150:Immunohistochemistry; D008833:Microcirculation; D011108:Polymers / Q000737:chemistry; D013499:Surface Properties; D013696:Temperature; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods*
D001672:Biocompatible Materials; D011108:Polymers