Determination of Electrical Stimuli Parameters To Transdifferentiate Genetically Engineered Mesenchymal Stem Cells into Neuronal or Glial Lineages

Abstract

This study investigated the effect of electrical stimuli parameters using graphene-based devices for the transdifferentiation of genetically engineered brain-derived neurotrophic factor (BDNF) hypersecreting mesenchymal stem cells (BDNF-MSCs) into neuronal or glial lineages. The results suggest that BDNF-MSCs have the tendency to transdifferentiate into both neuronal and Schwann cell (SC)-like phenotypes at lower voltages (25-50 mV). However, as the applied voltage changed from 25 to 100 mV at 50 Hz, the transdifferentiation of BDNF-MSCs yielded more into SC-like phenotypes and resulted in complete transdifferentiation into SC-like phenotypes at 100 mV and 50 Hz. With an increase in voltage to 100 mV, the complete transdifferentiation to SC-like phenotypes also resulted in enhanced paracrine activity leading to total secretion of nerve growth factor (NGF) up to 50 ng/mL with pronounced biological activity, causing neurite extension of 4 μm/cell on PC12-TrkB cells. Moreover, 90% of the transdifferentiated cells demonstrated significant myelination potential. The contact co-culture of BDNF-MSCs with adult hippocampal progenitor cells (AHPCs) in the presence of electrical stimuli resulted in differentiation of BDNF-MSCs into SC-like phenotypes accompanied by synergistic neurite extension of AHPCs. Overall, this study demonstrates the possibility of controlling simultaneous and spatial differentiation of MSCs into selected neuronal and glial lineages at desired ratios via changes in electrical stimuli through graphene-based devices and can contribute to the development of novel cell-based strategies for nervous system rescue and repair. Lay Summary This work evaluates the effect of different electrical stimuli conditions applied through inkjet-printed and laser-annealed graphene-based interdigitated circuits on the differentiation behavior of mesenchymal stem cells. Our results suggested that it is possible to spatially and locally control the differentiation of mesenchymal stem cells into final lineage type (glial or neuronal) by manipulating the electrical stimuli. The future work will include the control of stem cell differentiation and fate commitment in an in vivo model using electrical stimuli.

Ultrasonication effects on graphene composites in neural cell cultures

“…These assumptions find confirmation in the results of experiments by several researchers, including , Heo et al (2011), Uz et al (2020). The relevant literature review features reports on the effectiveness of weak electric field electrostimulation with voltage values ranging from 4.5 mV (Heo et al, 2011), 10 mV and 25, 50, and 100 mV (Das et al, 2017;Uz et al, 2020). However, in some cases, despite achieving an increased rate of cell count, electrostimulation only slightly affected the differentiation of cells on the graphene substrate and thus the formation of cell networks (Heo et al, 2011;.…”

“…In the study, the 5 V voltage amplitude was used. However, the literature reports confirm the efficiency of even lower stimulating voltages (Heo et al, 2011;Das et al, 2017;Huang and Zhu, 2019;Uz et al, 2020). This issue will be addressed in subsequent research.…”

“…In addition, the resistance of the culture substrate should be as low as possible to apply a signal with a lower electrical potential (Heo et al, 2011;Krukiewicz et al, 2019). These assumptions find confirmation in the results of experiments by several researchers, including , Heo et al (2011), Uz et al (2020). The relevant literature review features reports on the effectiveness of weak electric field electrostimulation with voltage values ranging from 4.5 mV (Heo et al, 2011), 10 mV and 25, 50, and 100 mV (Das et al, 2017;Uz et al, 2020).…”

“…Stimulation parameters such as period, frequency, and electrical voltage value need to be tailored to the specific cell group (Meng, 2014). Uz et al (2020) and Sherrell et al (2014) report different trends of cell differentiation depending on the stimulation voltage value. The authors observed a greater differentiation of cell populations into cells with a neural phenotype when stimulations with lower voltage values were applied.…”

“…The results of studies conducted by several other researchers support the hypothesis that electrical stimulation using graphene-based substrates and appropriate modulation of electrical parameters enhances the mechanisms of cell differentiation towards neural phenotypes. However, the low percentage of differentiated cells (10% of the population) remains a significant problem (Uz et al, 2020).…”