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Author Notes:

Correspondence: Jelena Vukasinovic ; Email: jvukasinovic@lenabio.com

This article was submitted to Neural Technology, a section of the journal Frontiers in Neuroscience

Present Address: Nathaniel J. Killian, Department of Neurosurgery, Massachusetts General Hospital, Boston, MA, USA

JV conceived the project. All authors designed the experiments. NK, VV, and JV performed the experiments. NK analyzed the data. NK and JV wrote the manuscript.

All authors edited the manuscript.

We thank Multi Channel Systems MCS GmbH for graciously providing pMEAs for this work.

We thank Michelle LaPlaca, Ph.D. and James Shoemaker for sharing cell culture equipment and brain tissue; Michelle Kuykendal for sharing fluorescent dyes and reagents; and Alex Calhoun, Ming-fai Fong, Jon Newman, and Riley Zeller-Townson for technical advice.

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. JV is the president and a shareholder of Lena Biosciences, Inc. The tool was developed by Lena Biosciences, Inc. Experiments were performed at Georgia Tech by NK through a sub-award, and at Lena Biosciences by VV who was employed by Lena Biosciences. VV holds no shares of Lena Biosciences, Inc.

Subjects:

Research Funding:

This work was supported by the National Institutes of Health SBIR awards to Lena Biosciences, Inc. 1 R43 NS065543 and 5 R43 NS065543 and 5 R01 NS079757 to Georgia Tech.

Keywords:

  • perforated microelectrode array
  • neurons
  • brain slice
  • three-dimensional culture
  • MEA

A Device for Long-Term Perfusion, Imaging, and Electrical Interfacing of Brain Tissue In vitro

Tools:

Journal Title:

Frontiers in Neuroscience

Volume:

Volume 10

Publisher:

, Pages 135-135

Type of Work:

Article | Final Publisher PDF

Abstract:

Distributed microelectrode array (MEA) recordings from consistent, viable, ≥500 μm thick tissue preparations over time periods from days to weeks may aid in studying a wide range of problems in neurobiology that require in vivo-like organotypic morphology. Existing tools for electrically interfacing with organotypic slices do not address necrosis that inevitably occurs within thick slices with limited diffusion of nutrients and gas, and limited removal of waste. We developed an integrated device that enables long-term maintenance of thick, functionally active, brain tissue models using interstitial perfusion and distributed recordings from thick sections of explanted tissue on a perforated multi-electrode array. This novel device allows for automated culturing, in situ imaging, and extracellular multi-electrode interfacing with brain slices, 3-D cell cultures, and potentially other tissue culture models. The device is economical, easy to assemble, and integrable with standard electrophysiology tools. We found that convective perfusion through the culture thickness provided a functional benefit to the preparations as firing rates were generally higher in perfused cultures compared to their respective unperfused controls. This work is a step toward the development of integrated tools for days-long experiments with more consistent, healthier, thicker, and functionally more active tissue cultures with built-in distributed electrophysiological recording and stimulation functionality. The results may be useful for the study of normal processes, pathological conditions, and drug screening strategies currently hindered by the limitations of acute (a few hours long) brain slice preparations.

Copyright information:

© 2016 Killian, Vernekar, Potter and Vukasinovic.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/).

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