The overall goal of this AREA renewal application is to develop microchip approaches that can be used to study cell-to-cell interactions at the molecular level. There are many examples in which the interaction of different cell types plays a role in normal biological function or in the onset of disease. One example is the interaction between neurons and glia cells that have undergone inflammation. It has been suggested that the degeneration of dopaminergic neurons that is prevalent in Parkinson's disease may be related to chronic inflammation of microglia, with the microglia producing nitric oxide and other reactive oxygen species that interact with the neurons. Another example is the vasodilatation process, where it has been shown that red blood cells, when exposed to hypoxic conditions or deformation, interact with endothelial cells via ATP release, leading to the production of nitric oxide and subsequent smooth muscle relaxation. Impairment of the ATP release from red blood cells leads to less nitric oxide production and has been postulated to play a role in several diseases. While there have been numerous examples of immobilizing cells on-chip and detecting a specific analyte released from the cells, there have been few reports of integrating multiple cell types on chip where cell-to-cell communication can be studied in a manner where numerous neurotransmitters/products can be monitored. In this grant we propose to continue the development of microchip approaches that enable the integration of cell immobilization with a general analysis step (electrophoresis), where a variety of analyses that are either released or transported through a layer of cells can be separated and subsequently detected via electrochemistry. Importantly, we propose to develop technology that will enable a researcher to monitor cell- to-cell communication between 2 layers of immobilized cells (PC 12 cells and microglia cells) or between a flowing stream of cells (red blood cells) and a layer of immobilized cells (endothelial cells). In addition, we propose to expand this microchip approach to enable in vivo studies by integrating microdialysis sampling, which can be used to study the interactions of different cell types by stereotaxically implanting a probe in the region of interest, with segmented flow, microchip electrophoresis, and electrochemical detection.
The specific aims of the grant are to 1) Use of reservoir-based cell immobilization and microchip electrophoresis with electrochemical detection to study the interaction between multiple cell types;2) Develop a microchip device that can be used to study cell-to-cell communication between red blood cells and an immobilized endothelium;and 3) Integration of microdialysis sampling and segmented flow with microchip electrophoresis and electrochemical detection for in vivo sampling.

Public Health Relevance

The overall goal of this AREA renewal application is to develop microchip approaches that can be used to study cell-to-cell interactions at the molecular level. The resulting technology will enable monitoring of cell-to- cell communication between 2 layers of immobilized cells or between flowing cells and an immobilized endothelium. The approaches will also enable in vivo studies by integrating microdialysis sampling with segmented flow, microchip electrophoresis, and electrochemical detection.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
2R15GM084470-03
Application #
8100044
Study Section
Instrumentation and Systems Development Study Section (ISD)
Program Officer
Edmonds, Charles G
Project Start
2004-12-10
Project End
2015-03-31
Budget Start
2011-04-01
Budget End
2015-03-31
Support Year
3
Fiscal Year
2011
Total Cost
$285,000
Indirect Cost
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
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
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
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
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
Pentecost, Amber M; Martin, R Scott (2015) Fabrication and Characterization of All-Polystyrene Microfluidic Devices with Integrated Electrodes and Tubing. Anal Methods 7:2968-2976
Bailey, Matthew R; Pentecost, Amber M; Selimovic, Asmira et al. (2015) Sheath-flow microfluidic approach for combined surface enhanced Raman scattering and electrochemical detection. Anal Chem 87:4347-55

Showing the most recent 10 out of 28 publications