Electrophysiological characterization of embryonic hippocampal neurons cultured in a 3D collagen hydrogel |
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| Authors: | Tao Xu Peter Molnar Cassie Gregory Mainak Das Thomas Boland James J. Hickman |
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| Affiliation: | 1. Department of Bioengineering, Clemson University, SC 29634, USA;2. NanoScience Technology Center, University of Central Florida, FL 32826, USA;3. Biomedical Science Center, University of Central Florida, FL 32826, USA;4. Department of Mechanical Engineering, University of Texas at El Paso, TX 79968, USA;1. Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Cheonan 330-714, South Korea;2. Biomaterials and Tissue Engineering Laboratory, Department of Nanobiomedical Science & WCU Research Center, Dankook University, Cheonan 330-714, South Korea;3. Glass Research Department, National Research Center, Cairo, Egypt;4. Department of Biomaterials Science, School of Dentistry, Dankook University, Cheonan 330-714, South Korea;1. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA;2. Faculty of Biomedical Engineering, Technion-Israel Institute of Technology, Technion City, Haifa 32000, Israel;1. Division of Neuroscience and Experimental Psychology, School of Biological Sciences, Faculty of Biology, Medicine and Health, The University of Manchester, Manchester Academic Health Science Centre, Manchester M13 9PL, UK;2. Division of Evolution and Genomic Sciences, School of Medical Sciences, Faculty of Biology, Medicine and Health, The University of Manchester, Manchester M13 9PL, UK;3. Manchester Institute of Biotechnology, The University of Manchester, Manchester M1 7DN, UK;1. Mechanobiology & Soft Matter Group, Laboratoire Interfaces et Fluides Complexes, Centre d’Innovation et de Recherche en Matériaux Polymères (CIRMAP), Research Institute for Biosciences, Université de Mons, 20, Place du Parc, B-7000 Mons, Belgium;2. Department of Neuroscience, University of Mons, Belgium;3. Laboratory for Soft Bioelectronics Interfaces, Centre for Neuroprosthetics, Ecole Fédérale Polytechnique de Lausanne, Switzerland |
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| Abstract: | Rat embryonic hippocampal neurons were cultured in (1) 3D collagen hydrogels as ‘entrapped’ evenly distributed cells, (2) at the interface of two collagen layers (sandwich model), and (3) on the surface of collagen coated coverslips (2D model). In the ‘entrapment’ model the neuronal processes grew out of the plane of the cell body and extended into the collagen matrix, in contrast to the sandwich model where the cells and their processes rarely left the plane in which they were seeded. Hippocampal neurons ‘entrapped’ in the 3D collagen gel grew the same number, but shorter, processes and exhibited improved survival compared to neurons cultured in the 2D model. There was no difference in the electrophysiological properties of the neurons cultured in the 3D compared to the 2D model except in the resting membrane potential and in the duration of the after-hyperpolarization. Spontaneous postsynaptic currents were recorded in 14- and 21-day-old 3D cultures evidencing functional synapse formation. Our results indicate that the physiological characteristics of 3D neuronal cultures are similar to traditional 2D cultures. However, functional 3D networks of hippocampal neurons will be necessary for multi-level circuit formation, which could be essential for understanding the basis of physiological learning and memory. |
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