Preparation of arrays of cell spheroids and spheroid-monolayer cocultures within a microfluidic device.
This study describes a novel method for generation of an array of three-dimensional (3D) multicellular spheroids within a microchannel in patterned cultures containing one or multiple cell types. This method uses a unique property of a cross-linked albumin coated surface in which the surface can be switched from non-adhesive to cell adhesive upon electrostatic adsorption of a polycation. Introduction of a solution containing albumin and a cross-linking agent into a microchannel with an array of microwells caused the entire surface, with the exception of the interior of the microwells, to become coated with the cross-linked albumin layer. Cells that were seeded within the microchannel did not adhere to the surface of the microchannel and became entrapped in the microwells. HepG2 cells seeded in the microwells formed 3D spheroids with controlled sizes and shapes depending upon the dimensions of the microwells. When the albumin coated surface was subsequently exposed to an aqueous solution containing poly(ethyleneimine) (PEI), adhesion of secondary cells, fibroblasts, occurred in the regions surrounding the arrayed spheroids. This coculture system can be coupled with spatially controlled fluids such as gradients and focused flow generators for various biological and tissue engineering applications.
Okuyama
Tomoaki
T
Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-0006, Japan.
Yamazoe
Hironori
H
Mochizuki
Naoto
N
Khademhosseini
Ali
A
Suzuki
Hiroaki
H
Fukuda
Junji
J
D000818:Animals; D002448:Cell Adhesion; D002449:Cell Aggregation; D002465:Cell Movement; D002470:Cell Survival; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D005456:Fluorescent Dyes; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D018874:Spheroids, Cellular / Q000166:cytology*; D041701:Swiss 3T3 Cells; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods
D005456:Fluorescent Dyes