Successful Differentiation of Neural Stem/Progenitor Cells Cultured on Electrically Adjustable Indium Tin Oxide (ITO) Surface

Abstract

In order to control differentiation of neural cells and guide the developed neurites to targets, polyelectrolyte multilayer (PEM) films were used because of their capability of modulation of electrical charged characteristics, thickness, and stiffness. In this work, we suggested that indium tin oxide (ITO) is an alternative surface to achieve the above-mentioned objectives. A microfluidic system with four culture chambers was developed and each chamber consisted of parallel ITO surfaces for the application of adjustable electrical field. Neural stem/progenitor cells (NSPCs) were respectively cultured on the ITO surfaces with and without PEM film, constructed by alternate adsorption of poly(L-lysine) (PLL) and poly(L-glutamic acid) (PLGA). Analyses of cell morphology, cytotoxicity, process outgrowth, differentiated cell types, and neuron functionality were compared between both surfaces. In this study, NSPCs successfully differentiated on ITO surface with electrical stimulation. The optimal electrical potential was found to be 80 mV that could stimulate the longest process, i.e., >300 μm, after 3 days culture. Cell differentiation, process development, and functionality of differentiated neuron on ITO surface were shown to be strongly controlled by the electrical stimulation that can be simply adjusted by external equipment. The electrically adjustable cell differentiation reported here could potentially be applied to neurochip for the study of neural signal transmission in a well-constructed network.

Nanostructured Multilayer Films Assembled from Poly(dopamine)‐Coated Carbon Nanotubes for Controlling Cell Behavior

“…[36] In our PC12 assay results, we found that PC12 cells can adhere on all three surfaces, PLL/ CNTs@PDA, PDA, and PLL surfaces (Figure 8), which is in accord with the previous references. [30,36] It is also observed that the PLL/CNTs@PDA multilayerf ilms favor the elongation of neural synapses compared with the PDA and PLL surfaces. This phenomenon was attributed to the influence of the nanotopographic structure of the PLL/CNTs@PDA multilayer films, which has been proved by other researches focusingo nt he relationship between electrospun nano-fibrous scaffolds and PC12 cell growth.…”

“…For instance, PLL‐grafted hydrogels greatly enhance PC12 cell survival in encapsulation, proliferation, and neurite growth . One research paper indicates that PDA can also promote adhesion and differentiation of PC12 cells . In our PC12 assay results, we found that PC12 cells can adhere on all three surfaces, PLL/CNTs@PDA, PDA, and PLL surfaces (Figure ), which is in accord with the previous references .…”

“…One research paper indicates that PDA can also promote adhesion and differentiation of PC12 cells . In our PC12 assay results, we found that PC12 cells can adhere on all three surfaces, PLL/CNTs@PDA, PDA, and PLL surfaces (Figure ), which is in accord with the previous references . It is also observed that the PLL/CNTs@PDA multilayer films favor the elongation of neural synapses compared with the PDA and PLL surfaces.…”

Construction of a graphene/polypyrrole composite electrode as an electrochemically controlled release system

“…54 Meanwhile, electrical stimulation mediates the stem cell adhesion, migration or differentiation behaviors. 12 For potential use in nerve tissue engineering, NPCs was selected to verify the biocompatibility of composite electrode in vitro. NPCs cultured on the substrates aer 5 days were stained with Calcein-AM and EthD-1, respectively.…”

“…9,10 Among these systems, the electro-responsive system is emerging to present a controlled release prole that mimics physiological processes in electroactive tissues owing to its intriguing properties of the versatility on the design of electrical stimulation devices or DDSs, 11 as well as the controllability through the tunable magnitude of voltage, frequency and pulse time. Besides, it has been reported that mild electrical stimulation facilitated the promotion of cell proliferation or differentiation, 12 providing the potential of electro-responsive DDSs in tissue engineering.…”