title: Creating two-dimensional patterned substrates for protein and cell confinement.
abstract: Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.(1) While microcontact printing can be employed to directly print a large number of molecules, including proteins,(2) DNA,(3) and silanes,(4) the formation of self-assembled monolayers (SAMs) from long chain alkane thiols on gold provides a simple way to confine proteins and cells to specific patterns containing adhesive and resistant regions. This confinement can be used to control cell morphology and is useful for examining a variety of questions in protein and cell biology. Here, we describe a general method for the creation of well-defined protein patterns for cellular studies.(5) This process involves three steps: the production of a patterned master using photolithography, the creation of a PDMS stamp, and microcontact printing of a gold-coated substrate. Once patterned, these cell culture substrates are capable of confining proteins and/or cells (primary cells or cell lines) to the pattern. The use of self-assembled monolayer chemistry allows for precise control over the patterned protein/cell adhesive regions and non-adhesive regions; this cannot be achieved using direct protein stamping. Hexadecanethiol, the long chain alkane thiol used in the microcontact printing step, produces a hydrophobic surface that readily adsorbs protein from solution. The glycol-terminated thiol, used for backfilling the non-printed regions of the substrate, creates a monolayer that is resistant to protein adsorption and therefore cell growth.(6) These thiol monomers produce highly structured monolayers that precisely define regions of the substrate that can support protein adsorption and cell growth. As a result, these substrates are useful for a wide variety of applications from the study of intercellular behavior(7) to the creation of microelectronics.(8) While other types of monolayer chemistry have been used for cell culture studies, including work from our group using trichlorosilanes to create patterns directly on glass substrates,(9) patterned monolayers formed from alkane thiols on gold are straight-forward to prepare. Moreover, the monomers used for monolayer preparation are commercially available, stable, and do not require storage or handling under inert atmosphere. Patterned substrates prepared from alkane thiols can also be recycled and reused several times, maintaining cell confinement.(10).
authors.affiliation: Department of Chemistry, Washington University in St. Louis, USA.
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Johnson
authors.forename: Natalie A
authors.initials: NA
authors.lastname: LaFranzo
authors.forename: Joshua A
authors.initials: JA
authors.lastname: Maurer
mesh_terms: D003584:Cytological Techniques / Q000379:methods*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods*; D009757:Nylons / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D004129:Dimethylpolysiloxanes; D009757:Nylons; D011506:Proteins; D013438:Sulfhydryl Compounds; C085707:poly(dimethylsiloxane)-polyamide copolymer; D006046:Gold