Thermally responsive polymer-grafted surfaces facilitate patterned cell seeding and co-culture

Tissue engineering constructs that effectively duplicate natural tissue fun ction must also maintain tissue architectural and organization features, pa rticularly the integration of multiple cell types preserving distinct, inte grated phenotypes. Cell-cell communication and biochemical cross-talk have been shown to be essential for the maintenance of differentiated cell funct ions in tissues and organs. Current limitations of cell-culture hinder prog ress in understanding the features and dynamics of heterotypic cell communi cation pathways critical to developing more sophisticated or effective tiss ue-engineered devices. We describe a method to conveniently electron-beam p attern cell culture surfaces with thermo-responsive polymer chemistry that exploits changes in cell-polymer adhesive interactions over a temperature w indow amenable for high-throughput cell culture. Cells seeded on these patt erned surfaces at 20 degreesC adhere only to surface areas lacking thermo-r esponsive grafting chemistry: grafted domains at 20 degreesC are hydrophili c and non-cell adhesive. The culture temperature is then increased to 37 de greesC, collapsing the hydrated grafted chemistry. A second cell type is ad ded to the culture and adheres only to these exposed relatively hydrophobic grafted patterns. Both cell types can then be effectively co-cultured at 3 7 degreesC under multiple conditions. Long-term cell pattern fidelity and d ifferentiated cell functions characteristic of each co-planar cell type are observed. This method is simple and has few limitations, compared with oth er existing co-culture methods. (C) 2001 Elsevier Science Ltd. All rights r eserved.