Preprint Brief Report Version 1 Preserved in Portico This version is not peer-reviewed

Screening Biophysical Sensors and Actuators that Influence Wound Healing in Human Induced Pluripotent Stem Cell-Derived Neurons

Version 1 : Received: 4 July 2022 / Approved: 6 July 2022 / Online: 6 July 2022 (03:52:01 CEST)

How to cite: Pai, V.; Cooper, B.; Levin, M. Screening Biophysical Sensors and Actuators that Influence Wound Healing in Human Induced Pluripotent Stem Cell-Derived Neurons. Preprints 2022, 2022070083 (doi: 10.20944/preprints202207.0083.v1). Pai, V.; Cooper, B.; Levin, M. Screening Biophysical Sensors and Actuators that Influence Wound Healing in Human Induced Pluripotent Stem Cell-Derived Neurons. Preprints 2022, 2022070083 (doi: 10.20944/preprints202207.0083.v1).

Abstract

All living cells maintain a charge distribution across their cell membrane (membrane potential) by carefully controlled ion fluxes. These bioelectric signals regulate cell behavior (such as migration, proliferation, differentiation) as well as higher-level tissue and organ patterning. Thus, voltage gradients represent an important parameter for diagnostics as well as a promising target for therapeutic interventions in birth defects, injury, and cancer. However, despite much progress in cell and molecular biology, little is known about bioelectric states in human stem cells. Here, we present simple methods to simultaneously track ion dynamics, membrane voltage, cell morphology, and cell activity (pH and ROS), using fluorescent reporter dyes in living human neurons derived from induced neural stem cells (hiNSC). We developed and tested functional protocols for manipulating ion fluxes, membrane potential, and cell activity, and tracking neural responses to wounding and re-innervation in vitro. Finally, using morphology sensor, we tested and quantified the ability of physiological actuators (neurotransmitters and pH) to manipulate nerve wound re-innervation. These methods are not specific to a particular cell type and should be broadly applicable to the study of bioelectrical controls across a wide range of combinations of models and endpoints.

Keywords

bioelectricity; ion flux; membrane potential; live sensor dyes; pH; serotonin; acetylcholine; GABA; hiNSC

Subject

BIOLOGY, Anatomy & Morphology

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