Generation of a patterned co-culture system composed of adherent cells and immobilized nonadherent cells

• Published in: Acta biomaterialia Vol. 31; pp. 231 - 240
• Main Authors: Yamazoe, Hironori, Ichikawa, Takashi, Hagihara, Yoshihisa, Iwasaki, Yasuhiko
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.02.2016
• Subjects: Adsorption; Albumins - chemistry; Animals; Biocompatible Materials; Caco-2 Cells; Cancer; Cancer microenvironment; Cell Adhesion; Cell Communication; Cell immobilization; Cell Line; Cell membrane anchoring reagent; Cells, Cultured; Cells, Immobilized; Circular Dichroism; Coculture Techniques - methods; Cross-Linking Reagents - chemistry; Culture; Drugs; Humans; Jurkat Cells; Mice; Microscopy, Phase-Contrast; Monocytes - cytology; Myeloid Cells - cytology; Neoplasms - drug therapy; Neoplasms - pathology; Neuroblastoma - metabolism; Patterned co-culture; Protein adsorption; Protein Structure, Tertiary; Reagents; Real-Time Polymerase Chain Reaction; Serum albumin; Surface Properties; Tissue engineering; Tissue Engineering - instrumentation; Tissue Engineering - methods; Utilities; Serum albumin; Cell membrane anchoring reagent; Protein adsorption; Patterned co-culture; Cancer microenvironment; Cell immobilization
• ISSN: 1742-7061; 1878-7568
• DOI: 10.1016/j.actbio.2015.12.016

[Display omitted] Patterned co-culture is a promising technique used for fundamental investigation of cell–cell communication and tissue engineering approaches. However, conventional methods are inapplicable to nonadherent cells. In this study, we aimed to establish a patterned co-culture system composed of adherent and nonadherent cells. Nonadherent cells were immobilized on a substrate using a cell membrane anchoring reagent conjugated to a protein, in order to incorporate them into the co-culture system. Cross-linked albumin film, which has unique surface properties capable of regulating protein adsorption, was used to control their spatial localization. The utility of our approach was demonstrated through the fabrication of a patterned co-culture consisting of micropatterned neuroblastoma cells surrounded by immobilized myeloid cells. Furthermore, we also created a co-culture system composed of cancer cells and immobilized monocytes. We observed that monocytes enhanced the drug sensitivity of cancer cells and its influence was limited to cancer cells located near the monocytes. Therefore, the incorporation of nonadherent cells into a patterned co-culture system is useful for creating culture systems containing immune cells, as well as investigating the influence of these immune cells on cancer drug sensitivity. Various methods have been proposed for creating patterned co-culture systems, in which multiple cell types are attached to a substrate with a desired pattern. However, conventional methods, including our previous report published in Acta Biomaterialia (2010, 6, 526–533), are unsuitable for nonadherent cells. Here, we developed a novel method that incorporates nonadherent cells into the co-culture system, which allows us to precisely manipulate and study microenvironments containing nonadherent and adherent cells. Using this technique, we demonstrated that monocytes (nonadherent cells) could enhance the drug sensitivity of cancer cells and that their influence had a limited effective range. Thus, our technique is useful for recreating complex tissues in order to investigate cellular interactions involving nonadherent cells.