The overall goal of this AREA renewal application is focused on the development of new microchip approaches to study cell-to-cell-interactions. Cell-to-cell communication is an important component in understanding cell function. A better understanding of these interactions could lead to important advances in the medical and neuroscience fields, but a lack of technology currently limits the ability to study the interaction between cell lines at the molecular level.
In aim #1, we propose to develop a new and innovative chip-to-chip approach that can be used to study the interaction between two adherent cells, PC 12 cells and microglia cells. While there have been reports of using highly integrated microchip devices for analyzing cells on-chip, they are relatively low throughput and if any part of the chip fails the entire experiment is sacrificed. To address these issues, we propose in aim #1 the development of a new approach, a polystyrene (PS)-based chip-to-chip device in which a cell culture/immobilization device is isolated from the analysis device. This more modular approach will enable the numerous advantages of doing cell culture in microchip devices and, when analysis is desired by microchip electrophoresis and electrochemical detection, our recently reported ability to encapsulate both tubing and electrodes in PS devices will lead to a low dead volume connection to direct flow from the cell chip to the analysis chip. In addition, we propose a new type of device that interfaces a bilayer PDMS-based microchip with an embedded fused-silica capillary to enhance the separation capabilities. In essence, this approach will bridge the fundamental advantages of PDMS devices (including the ability to incorporate valves and electrochemical detection) with widely used fused-silica capillary for more efficient separations.
In specific aim #2, we propose a new type of planar membrane device that can be used to study the interaction between a flowing cell line (red blood cells) and adherent cells (endothelial cells). The focus of this aim is detection of one of the key chemical messengers, nitric oxide (NO), an analytical challenging molecule that has a very short half-life in vivo. The development of a new and innovative system capable of electrochemically monitoring NO release from immobilized endothelial cells as a response to RBC derived ATP release within seconds is essential to a better understanding of cellular signaling at the molecular level and could lead to further knowledge in the cardiovascular field. In this aim, we propose a novel PS-based device that uses a pillar array to create a planar membrane that will allow small molecules such as NO to pass through the pores created by the array into an adjacent channel (collector channel), where they will be electrochemically detected using our recently reported Pt-black electrodes. The planar membrane approach will enable visualization of all cells lines as well as allowing the collector channel and the detection electrodes to be aligned in very close proximity to the membrane (but away from the channel containing cells), which is essential for detecting a short-lived molecule such as NO.

Public Health Relevance

The overall goal of this AREA renewal application is focused on the development of new microchip approaches to study cell-to-cell-interactions. A better understanding of cell-to-cell interactions could lead to important advances in the medical and neuroscience fields, but a lack of technology currently limits the ability to study the interaction between cell lines at the molecular level. We propose the development of a new 'chip-to-chip' approach that can be used to study the interaction between 2 adherent cells (PC 12 cells, a commonly used neuronal cell model, and microglia cells) as well as a new type of planar membrane device that can be used to study the interaction between a flowing cell line (red blood cells) and adherent cells (endothelial cells).

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
2R15GM084470-04A1
Application #
8874579
Study Section
Special Emphasis Panel (ZRG1-BST-F (81))
Program Officer
Edmonds, Charles G
Project Start
2004-12-10
Project End
2018-08-31
Budget Start
2015-09-30
Budget End
2018-08-31
Support Year
4
Fiscal Year
2015
Total Cost
$303,000
Indirect Cost
$103,000
Name
Saint Louis University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
050220722
City
Saint Louis
State
MO
Country
United States
Zip Code
63103
Mehl, Benjamin T; Martin, R Scott (2018) Enhanced Microchip Electrophoresis Separations Combined with Electrochemical Detection Utilizing a Capillary Embedded in Polystyrene. Anal Methods 10:37-45
Chen, Chengpeng; Townsend, Alexandra D; Hayter, Elizabeth A et al. (2018) Insert-based microfluidics for 3D cell culture with analysis. Anal Bioanal Chem 410:3025-3035
Chen, Chengpeng; Townsend, Alexandra D; Sell, Scott A et al. (2017) Microchip-based 3D-Cell Culture Using Polymer Nanofibers Generated by Solution Blow Spinning. Anal Methods 9:3274-3283
Bailey, Matthew R; Martin, R Scott; Schultz, Zachary D (2016) Role of Surface Adsorption in the Surface-Enhanced Raman Scattering and Electrochemical Detection of Neurotransmitters. J Phys Chem C Nanomater Interfaces 120:20624-20633
Forzano, Anna V; Becirovic, Vedada; Martin, R Scott et al. (2016) Integrated Electrodes and Electrospray Emitter for Polymer Microfluidic Nanospray-MS Interface. Anal Methods 8:5152-5157
Munshi, Akash S; Martin, R Scott (2016) Microchip-based electrochemical detection using a 3-D printed wall-jet electrode device. Analyst 141:862-9
Chen, Chengpeng; Mehl, Benjamin T; Munshi, Akash S et al. (2016) 3D-printed Microfluidic Devices: Fabrication, Advantages and Limitations-a Mini Review. Anal Methods 8:6005-6012
Townsend, Alexandra D; Wilken, Gerald H; Mitchell, Kyle K et al. (2016) Simultaneous analysis of vascular norepinephrine and ATP release using an integrated microfluidic system. J Neurosci Methods 266:68-77
Chen, Chengpeng; Mehl, Benjamin T; Sell, Scott A et al. (2016) Use of electrospinning and dynamic air focusing to create three-dimensional cell culture scaffolds in microfluidic devices. Analyst 141:5311-20
Johnson, Alicia S; Mehl, Benjamin T; Martin, R Scott (2015) Integrated hybrid polystyrene-polydimethylsiloxane device for monitoring cellular release with microchip electrophoresis and electrochemical detection. Anal Methods 7:884-893

Showing the most recent 10 out of 30 publications