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Hindi Research Article
Novel Microchip Technique for the Transfer of Spheroids as Floating Cultures to Micropatterned-Adherent Cultures
| Sakai Y1,2 and Nakazawa K1* | |
| 1Department of Life and Environment Engineering, The University of Kitakyushu, Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan | |
| 2Research Fellow of the Japan Society for the Promotion of Science (JSPS), Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan | |
| Corresponding Author : | Kohji Nakazawa Department of Life and Environment Engineering The University of Kitakyushu, Hibikino Wakamatsu-ku, Kitakyushu Fukuoka 808-0135, Japan Tel: +81-93-695-3292 Fax: +81-93-695-3359 E-mail: nakazawa@kitakyu-u.ac.jp |
| Received June 18, 2010; Accepted December 05, 2011; Published December 10, 2011 | |
| Citation: Sakai Y, Nakazawa K (2011) Novel Microchip Technique for the Transfer of Spheroids as Floating Cultures to Micropatterned-Adherent Cultures. J Biochip Tissue chip S4:001. doi:10.4172/2153-0777.S4-001 | |
| Copyright: © 2011 Sakai Y, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. | |
Abstract
Spherical multicellular aggregate (spheroid) culture has attracted attention as useful technique for tissue engineering or regenerative medicine research and cell-based assay studies. Although spheroids are generated using floating culture method on non-adhesive surface, the transfer of floating spheroids to adherent culture holds importance for various applications. In this paper, we successfully established a novel microchip technique which allowed the formation of spheroids with uniform size and their transfer from a floating condition to a micropatterned adherent culture. A spheroid transfer chip (ST chip) contained 270 microwells (600 ?m in diameter, 600 ?m in depth, and 660 ?m in pitch) composed of a through-hole PMMA frame and a PDMS sheet, and the surface of microwells was modified with PEG to create non-adhesive surface. Mouse ES cells, NIH 3T3 cells, HepG2 cells, and rat primary hepatocytes spontaneously formed spheroids in each microwell of the chips. The micropatterned adherent culture of spheroids was realized by simple procedures, which the ST chip was flipped onto a collagencoated dish, and then the PDMS sheet was peeled off after the spheroids fell onto the dish. The transferred spheroids gradually spread on the collagen-coated dish, and the overlap of cells extending from each spheroid occurred in order of mouse ES cells, 3T3 cells, HepG2 cells, and primary rat hepatocytes. This difference may be due to cell proliferation ability, cell viability in the spheroid, and spheroid size. Because the chip characteristics such as the microwell size, the pitch between spheroids, spheroid sizes, and spheroid number can be designed according to the experimental needs, this chip technique is a promising tool for spheroid study.
