title: Cell co-culture patterning using aqueous two-phase systems.
abstract: Cell patterning technologies that are fast, easy to use and affordable will be required for the future development of high throughput cell assays, platforms for studying cell-cell interactions and tissue engineered systems. This detailed protocol describes a method for generating co-cultures of cells using biocompatible solutions of dextran (DEX) and polyethylene glycol (PEG) that phase-separate when combined above threshold concentrations. Cells can be patterned in a variety of configurations using this method. Cell exclusion patterning can be performed by printing droplets of DEX on a substrate and covering them with a solution of PEG containing cells. The interfacial tension formed between the two polymer solutions causes cells to fall around the outside of the DEX droplet and form a circular clearing that can be used for migration assays. Cell islands can be patterned by dispensing a cell-rich DEX phase into a PEG solution or by covering the DEX droplet with a solution of PEG. Co-cultures can be formed directly by combining cell exclusion with DEX island patterning. These methods are compatible with a variety of liquid handling approaches, including manual micropipetting, and can be used with virtually any adherent cell type.
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Michigan, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Frampton
authors.forename: Joshua B
authors.initials: JB
authors.lastname: White
authors.forename: Abin T
authors.initials: AT
authors.lastname: Abraham
authors.forename: Shuichi
authors.initials: S
authors.lastname: Takayama
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018920:Coculture Techniques / Q000379:methods*; D003911:Dextrans / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology; D006367:HeLa Cells; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D044367:Phase Transition; D011092:Polyethylene Glycols / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D003911:Dextrans; D014867:Water; D011092:Polyethylene Glycols