Stepwise assembly of micropatterned co-cultures using photoresponsive culture surfaces and its application to hepatic tissue arrays

• Published in: Biotechnology and bioengineering Vol. 103; no. 3; pp. 552 - 561
• Main Authors: Kikuchi, Kyoko, Sumaru, Kimio, Edahiro, Jun-Ichi, Ooshima, Yuki, Sugiura, Shinji, Takagi, Toshiyuki, Kanamori, Toshiyuki
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 15.06.2009; Wiley; Wiley Subscription Services, Inc
• Subjects: Animals; Biological and medical sciences; Biomedical research; Biotechnology; Cell Adhesion - radiation effects; Cell culture; Cell Line; Coculture Techniques; Cytochrome P-450 CYP3A - biosynthesis; Fibroblasts; Fundamental and applied biological sciences. Psychology; Gene Expression Profiling; Hepatocytes; HepG2; Humans; Liver; Mice; P450 expression; photoresponsive culture surface; stepwise cell patterning; tissue array; Tissue Culture Techniques - methods; Tissue engineering; Tissue Engineering - methods; Tissues; Ultraviolet Rays; Array; tissue array; Tissue; Patterning; Cytochrome P450; Liver; P450 expression; HepG2; stepwise cell patterning; photoresponsive culture surface; Application; Culture
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.22253

Cell micropatterning, a method to place cells at arbitrary regions, is becoming an essential tool to conduct cell biology and tissue engineering. Conventional cell patterning techniques usually allow only single patterning with single cell type on the same culture surface. However, biomedical research today requires even sophisticated fabrication methods that require spatiotemporal control of multiple cell arrangements. Here we introduce in situ cell micropatterning system which enables stepwise cell patterning using a photoresponsive cell culture surface (PRCS) whose cell adhesiveness could be altered by the UV irradiation. To demonstrate an application to tissue engineering, a liver‐mimic tissue array was fabricated and liver‐specific gene expressions were quantified with real time PCR. Patterned co‐culture systems composed of HepG2 spheroids with Balb/3T3 were fabricated, and the optimum spheroid diameter, which yielded the highest cellular functions, was determined to be 150 µm. After 20 days of patterned co‐culture of HepG2 spheroids and Balb/3T3, CYP3A4 expression increased 50‐fold higher than conventionally cultured HepG2; CYP3A4 expression was 20% higher than randomly co‐cultured HepG2 and Balb/3T3. Thus the combination of PRCS and the photomask‐free irradiation apparatus showed the versatility of experimental setups and proved to be a powerful tool for biomedical studies. Biotechnol. Bioeng. 2009;103: 552–561. © 2009 Wiley Periodicals, Inc.