alexa Effects of Culture Conditions on a Micropatterned Co-cu
ISSN: 2155-9821

Journal of Bioprocessing & Biotechniques
Open Access

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Research Article

Effects of Culture Conditions on a Micropatterned Co-culture of Rat Hepatocytes with 3T3 cells

Kohji Nakazawa*, Yukako Shinmura, Ami Higuchi and Yusuke Sakai
Department of Life and Environment Engineering, The University of Kitakyushu 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan
Corresponding Author : Dr. Kohji Nakazawa
Department of Life and Environment Engineering
The University of Kitakyushu
1-1 Hibikino, Wakamatsu-ku
Kitakyushu, Fukuoka 808-0135, Japan
Tel: +81-93-695-3292
Fax: +81-93-695-3359
E-mail: [email protected]
Received August 28, 2011; Accepted September 28, 2011; Published September 30, 2011
Citation: Nakazawa K, Shinmura Y, Higuchi A, Sakai Y (2011) Effects of Culture Conditions on a Micropatterned Co-culture of Rat Hepatocytes with 3T3 cells. J Bioprocess Biotechniq S3:002. doi: 10.4172/2155-9821.S3-002
Copyright: © 2011 Nakazawa K, 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.


We investigated the effect of culture conditions on the micropatterned co-culture of rat hepatocytes with 3T3 cells. A micropatterned chip was prepared using polydimethylsiloxane (PDMS) microstencil such that the chip contained 724 hepatocyte islands, each 500 μm in diameter, in a triangular arrangement with 800-μm pitch, in which hepatocytes were co-cultured with 3T3 cells. The hepatocytes in micropatterned co-culture exhibited hepatocellular morphology, and the micropatterned configuration of hepatocyte islands was maintained for several weeks of culture by supporting the heterotypic interface between the hepatocytes and 3T3 cells. The albumin secretion activity of hepatocytes was highest in the micropatterned co-culture but decreased in the random co-culture, micropatterned mono-culture (hepatocytes only), and random mono-culture (hepatocytes only) in that order. Furthermore, earlier formation of co-culture promoted higher functional activity of hepatocytes as compared to later formation, and hepatocyte functions were induced with an increasing the density of inoculated 3T3 cells. These results suggest that the formation of a heterotypic interaction at an early stage is important for maintaining high levels of hepatocyte functions. The findings of this study will provide information useful for designing co-culture conditions for liver tissue engineering and pharmacological and toxicological studies.

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