On-line observation of cell growth in a three-dimensional matrix on surface-modified microelectrode arrays

Abstract

Despite many successful applications of microelectrode arrays (MEAs), typical two-dimensional in-vitro cultures do not project the full scale of the cell growth environment in the three-dimensional (3D) in-vivo setting. This study aims to on-line monitor in-vitro cell growth in a 3D matrix on the surface-modified MEAs with a dynamic perfusion culture system. A 3D matrix consisting of poly(ethylene glycol) hydrogel supplemented with poly-D-lysine was subsequently synthesized in-situ on the self-assembled monolayers modified MEAs. FTIR spectrum analysis revealed a peak at 2100 cm−1 due to the degradation of the structure of the 3D matrix. After 2 wks, microscopic examination revealed that the non-degraded area was around 1500 μm2 and provided enough space for cell growth. Fluorescence microscopy revealed that the degraded 3D matrix was non-cytotoxic allowing the growth of NIH3T3 fibroblasts and cortical neurons in vitro. Time-course changes of total impedance including resistance and reactance were recorded for 8 days to evaluate the cell growth in the 3D matrix on the MEA. A consistent trend reflecting changes of reactance and total impedance was observed. These in-vitro assays demonstrate that our 3D matrix can construct a biomimetic system for cell growth and analysis of cell surface interactions.

Investigation into the Effect of Varied Functional Biointerfaces on Silicon Nanowire MOSFETs

“…The biointerface was created by immersing APTMS or MPTMS modified NW-MOSFETs in 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) citrate buffered solution of antibody against prostate-specific antigen (anti-PSA, Ortho-Clinical Diagnostics Johnson & Johnson, 34 kDa). The EDC-NHS was utilized to only activate terminal carboxyl (-COOH) group of antibody [16,20] and maintain exposure of its active site for antigen binding.…”

“…Surface modification techniques have been applied to functionalize FETs for the specific detection of ions [4,12], DNA [7,13], proteins [4,8], and viruses [14]. In addition, the surface modification or functionalization introduces functional groups and then creates an interface between the biological system and the electronic device for detection of neuronal signals [15] and observation of cell growth [16]. Organosilane molecules are popular candidates for reacting with hydroxyl-terminated surfaces [17].…”

Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

“…Figure 5A shows an electrochemical biosensor that incorporates the 3D culture in a synthetic hydrogel on a flat electrode surface. 85 Figure 5B shows the transduction principle and the resulting signals of the 2D and 3D culture-based electrochemical biosensors (Fig. 5B).…”

“…Lin et al used a flat microelectrode array to monitor the impedance change during the growth of NIH 3T3 fibroblasts and cortical neurons in a 3D matrix in a label-free manner. 85 Jeong et al reported a similar design of an electrochemical biosensor using several types of sol-gels (alginate, collagen, Matrigel) to grow A549 lung cancer cells in 3D on flat electrodes. 78 Cellular responses to anticancer drugs were successfully monitored.…”

From Cellular Cultures to Cellular Spheroids: Is Impedance Spectroscopy a Viable Tool for Monitoring Multicellular Spheroid (MCS) Drug Models?

“…Using a three chamber microfluidic device, it was found that when CAF were cultured adjacent to ACC-M, ACC-M cells invaded neighboring matrix and was reduced when MMP inhibitor was GM 6001 was introduced. Similar studies have proven EIS capable of monitoring growth rate of cells embedded inside of a 3-D matrix [119]. Impedance based sensors integrated within culture chambers could actively sense changes in the density of cells trapped within the matrix allowing for real-time cellular invasion assays, allowing for fully automated real-time monitoring of MCTS invasion.…”