title: Micropatterned culture of HepG2 spheroids using microwell chip with honeycomb-patterned polymer film.
abstract: Microwell chip culture is a promising technique for the generation of homogenous spheroids. We investigated the relationship between the structure of the bottom surface of microwell chip and the properties of HepG2 spheroid. We developed a microwell chip, the bottom surface of which consisted of a honeycomb-patterned polymer film (honeycomb film) that had a regular porous structure (HF chip). The chip comprised 270 circular microwells; each microwell was 600 μm in diameter and 600 μm in depth. At the center of the honeycomb film, an area, 200 μm in diameter, was modified with collagen to facilitate cell adhesion. With the exception of the collagen-coated area, the entire microwell was modified with polyethylene glycol to eliminate cell adhesion. HepG2 cells formed uniform spheroids when cultured in the microwells of HF chip. Furthermore, the cells passed through the porous structure of honeycomb film and formed spheroids at its opposite side. The spheroid growth of HepG2 cells cultured in HF chip was greater than that when the cells were culture in a microwell chip, the bottom surface of which was made of poly-methylmethacrylate (PMMA chip). The albumin secretion activity of HepG2 spheroids in HF chip was equal to that in PMMA chip. These results indicate that the microwell bottom with a porous structure enhances the cell growth and maintains well the spheroid function. Thus, HF chip is a promising platform for spheroid cell culture.
authors.affiliation: Production Engineering & Development Center Research & Development Management Headquarters, Fujifilm Corporation, 210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan; Institute of Multidisciplinary Research for Advanced Materials (IMRAM), Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Yamazaki
authors.affiliation: Production Engineering & Development Center Research & Development Management Headquarters, Fujifilm Corporation, 210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan.
authors.forename: Shun
authors.initials: S
authors.lastname: Gotou
authors.affiliation: Production Engineering & Development Center Research & Development Management Headquarters, Fujifilm Corporation, 210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan.
authors.forename: Koju
authors.initials: K
authors.lastname: Ito
authors.affiliation: Production Engineering & Development Center Research & Development Management Headquarters, Fujifilm Corporation, 210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan.
authors.forename: Souichi
authors.initials: S
authors.lastname: Kohashi
authors.affiliation: Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Goto
authors.affiliation: Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yoshiura
authors.affiliation: Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sakai
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials (IMRAM), Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yabu
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials (IMRAM), Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577, Japan; WPI-Advanced Institute for Materials Research, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), 4-1-8 Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Masatsugu
authors.initials: M
authors.lastname: Shimomura
authors.affiliation: Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan. Electronic address: nakazawa@kitakyu-u.ac.jp.
authors.forename: Kohji
authors.initials: K
authors.lastname: Nakazawa
mesh_terms: D000418:Albumins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000737:chemistry; D056945:Hep G2 Cells; D006801:Humans; D046208:Microchip Analytical Procedures; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology; D019904:Polymethyl Methacrylate / Q000737:chemistry / Q000494:pharmacology; D016062:Porosity; D018874:Spheroids, Cellular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
chemical_list: D000418:Albumins; D011092:Polyethylene Glycols; D003094:Collagen; D019904:Polymethyl Methacrylate
keywords: Cell proliferation; Collagen; HepG2 cells; Honeycomb-patterned polymer film; Microwell chip; Polyethylene glycol; Spheroid