title: Micropatterning neuronal cells on polyelectrolyte multilayers.
abstract: This paper describes an approach to adhere retinal cells on micropatterned polyelectrolyte multilayer (PEM) lines adsorbed on poly(dimethylsiloxane) (PDMS) surfaces using microfluidic networks. PEMs were patterned on flat, oxidized PDMS surfaces by sequentially flowing polyions through a microchannel network that was placed in contact with the PDMS surface. Polyethyleneimine (PEI) and poly(allylamine hydrochloride) (PAH) were the polyions used as the top layer cellular adhesion material. The microfluidic network was lifted off after the patterning was completed and retinal cells were seeded on the PEM/PDMS surfaces. The traditional practice of using blocking agents to prevent the adhesion of cells on unpatterned areas was avoided by allowing the PDMS surface to return to its uncharged state after the patterning was completed. The adhesion of rat retinal cells on the patterned PEMs was observed 5 h after seeding. Cell viability and morphology on the patterned PEMs were assayed. These materials proved to be nontoxic to the cells used in this study regardless of the number of stacked PEM layers. Phalloidin staining of the cytoskeleton revealed no apparent morphological differences in retinal cells compared with those plated on polystyrene or the larger regions of PEI and PAH; however, cells were relatively more elongated when cultured on the PEM lines. Cell-to-cell communication between cells on adjacent PEM lines was observed as interconnecting tubes containing actin that were a few hundred nanometers in diameter and up to 55 microm in length. This approach provides a simple, fast, and inexpensive method of patterning cells onto micrometer-scale features.
authors.affiliation: Semiconductor Electronics Division, National Institute of Standards and Technology, 100 Bureau Drive, MS 8120, Gaithersburg, Maryland 20899-8120, USA. darwin.reyes@nist.gov
authors.forename: Darwin R
authors.initials: DR
authors.lastname: Reyes
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Perruccio
authors.forename: S Patricia
authors.initials: SP
authors.lastname: Becerra
authors.forename: Laurie E
authors.initials: LE
authors.lastname: Locascio
authors.forename: Michael
authors.initials: M
authors.lastname: Gaitan
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D008422:Materials Testing; D044085:Microfluidics; D009474:Neurons / Q000166:cytology*; D011073:Polyamines / Q000737:chemistry*; D000071228:Polyelectrolytes; D051381:Rats; D012160:Retina / Q000166:cytology; D012828:Silicones / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D004129:Dimethylpolysiloxanes; D011073:Polyamines; D000071228:Polyelectrolytes; D012828:Silicones; C013830:baysilon