title: Effect of microwell chip structure on cell microsphere production of various animal cells.
abstract: The formation of three-dimensional cell microspheres such as spheroids, embryoid bodies, and neurospheres has attracted attention as a useful culture technique. In this study, we investigated a technique for effective cell microsphere production by using specially prepared microchip. The basic chip design was a multimicrowell structure in triangular arrangement within a 100-mm(2) region in the center of a polymethylmethacrylate (PMMA) plate (24x24 mm(2)), the surface of which was modified with polyethylene glycol (PEG) to render it nonadhesive to cells. We also designed six similar chips with microwell diameters of 200, 300, 400, 600, 800, and 1000 microm to investigate the effect of the microwell diameter on the cell microsphere diameter. Rat hepatocytes, HepG2 cells, mouse embryonic stem (ES) cells, and mouse neural progenitor/stem (NPS) cells formed hepatocyte spheroids, HepG2 spheroids, embryoid bodies, and neurospheres, respectively, in the microwells within 5 days of culture. For all the cells, a single microsphere was formed in each microwell under all the chip conditions, and such microsphere configurations remained throughout the culture period. Furthermore, the microsphere diameters of each type of cell were strongly positively correlated with the microwell diameters of the chips, suggesting that microsphere diameter can be factitiously controlled by using different chip conditions. Thus, this chip technique is a promising cellular platform for tissue engineering or regenerative medicine research, pharmacological and toxicological studies, and fundamental studies in cell biology.
authors.affiliation: Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu 808-0135, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sakai
authors.forename: Shirou
authors.initials: S
authors.lastname: Yoshida
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yoshiura
authors.forename: Rhuhei
authors.initials: R
authors.lastname: Mori
authors.forename: Tomoko
authors.initials: T
authors.lastname: Tamura
authors.forename: Kanji
authors.initials: K
authors.lastname: Yahiro
authors.forename: Hideki
authors.initials: H
authors.lastname: Mori
authors.forename: Yonehiro
authors.initials: Y
authors.lastname: Kanemura
authors.forename: Mami
authors.initials: M
authors.lastname: Yamasaki
authors.forename: Kohji
authors.initials: K
authors.lastname: Nakazawa
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation*; D049108:Cell Enlargement; D049109:Cell Proliferation; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D051379:Mice; D008904:Miniaturization; D051381:Rats; D018874:Spheroids, Cellular / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000295:instrumentation*