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https://hdl.handle.net/2142/82532
Description
Title
Cell Patterning: Building Living Neural Networks
Author(s)
Corey, Joseph Michael
Issue Date
1997
Doctoral Committee Chair(s)
Wheeler, Bruce C.
Department of Study
Neuroscience
Discipline
Neuroscience
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Engineering, Biomedical
Language
eng
Abstract
Neuronal growth can be controlled in vitro by plating cells at low density and by differential adhesion between the cell and substrate. This principle was used to develop a technology to pattern neurons in reduced networks consistent with recording from extracellular electrode arrays. The goals were to localize cell bodies (somata) to predefined areas and limit connectivity by reducing neurite growth to paths between the somata. Primary cultures of rat hippocampal neurons and B104 neuroblastoma cells were grown in serum-free culture on patterns fabricated on glass coverslips using several microlithographic methods. In three of the studies, adhesive grids having varying pathwidths, pathlengths (distances between intersections), and nodal (intersection) diameters were fabricated against less adhesive backgrounds. Not only did somata strongly prefer the adhesive patterns, but they also migrated to loci where the local area of adhesive material was higher. Neurons grown on laser-ablated polylysine grids exhibited best patterning (compliance) on grids with 80 $\mu$m pathlengths, moderate pattern compliance on 120 $\mu$m grids, and almost no compliance on 160 $\mu$m grids. The greatest compliance to pattern was 94%. Neurons grown on aminoalkane patterns against phenylsilane backgrounds exhibited a similar maximal compliance, improved compliance on 120 $\mu$m and 160 $\mu$m pathlengths, and improved compliance of neurites to paths, with an average of 77% of background squares free of neurites or cells connected to the pattern. B104 neuroblastomas cells, which patterned better than N1E-115, NG108, and B103 cell lines, were developed as a rapid assay to test patterned substrates. Primary hippocampal neurons were also grown on alternating stripes of polylysine versus stripes of extracellular matrix protein (laminin or pleiotrophin with or without polylysine). These substrates were fabricated by sequentially binding biomolecules to substrates with polydimethylsiloxane microstamps. On laminin/polylysine versus polylysine, 80% of neurons near the border extended their axons along the laminin/polylysine stripe or along the border between the two stripes. These cells also developed 2.5 times as many dendrites on the polylysine than on the laminin/polylysine. This technology should lead to the construction of neuronal circuits to investigate synaptic plasticity in vitro and tissue engineering and repair in vivo.
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