Electrical conditioning of adipose‐derived stem cells in a multi‐chamber culture platform

Abstract

This is the peer reviewed version of the following article A. Pavesi, M. Soncini, A. Zamperone, S. Pietronave, E. Medico, A. Redaelli, M. Prat and G. B. Fiore. Electrical conditioning of adipose-derived stem cells in a multi-chamber culture platform. Biotechnology and Bioengineering Volume 111, Issue 7, July 2014, Pages: 1452-1463 DOI: 10.1002 Abstract:In tissue engineering, several factors play key roles in providing adequate stimuli for cells differentiation, in particular biochemical and physical stimuli, which try to mimic the physiological microenvironments.Since electrical stimuli are important in the developing heart, we have developed an easy-to-use, costeffective cell culture platform, able to provide controlled electrical stimulation aimed at investigating the influence of the electric field in the stem cell differentiation process. This bioreactor consists of an electrical stimulator and twelve independent, petri-like culture chambers and a 3-D computational model was used to characterize the distribution and the intensity of the electric field generated in the cell culture volume. We explored the effects of monophasic and biphasic square wave pulse stimulation on a mouse adipose-derived stem cell line (m17.ASC) comparing cell viability, proliferation, protein and gene expression. Both monophasic (8V, 2ms, 1Hz) and biphasic (+4V, 1ms and -4V, 1ms; 1Hz) stimulation were compatible with cell survival and proliferation. Biphasic stimulation induced the expression of Connexin 43, which was found to localize also at the cell membrane, which is its recognized functional mediating intercellular electrical coupling. Electrically stimulated cells showed an induced transcriptional profile more closely related to that of neonatal cadiomyocytes, particularly for biphasic stimulation. The developed platform thus allowed to set-up precise conditions to drive adult stem cells toward a myocardial phenotype solely by physical stimuli, in the absence of exogenously added expensive bioactive molecules, and can thus represent a valuable tool for translational applications for heart tissue engineering and regeneration.

Electrical stimulation to optimize cardioprotective exosomes from cardiac stem cells

“…Kim et al first used EleS to pre-condition cells for the purpose of increasing the survival of transplanted CSC and enhancing their benefit to the ischemic heart (20). However, EleS has been used to promote the differentiation of CSC (23) and adipose-derived stem cells (24) into cardiomyocyte-type cells. Primarily, the difference between electrically pre-conditioning stem cells and inducing their differentiation may depend on the duration of EleS, as pre-conditioning was elicited by short-term EleS (3 hours) (20), while differentiation required long-term stimulation (14 days) (23).…”

Electrical and mechanical stimulation of cardiac cells and tissue constructs

“…Stimulated hiPSC biowire constructs showed increased myofibril ultra-structural organization (demonstrated by α-actinin and cTnT staining and imaging of desmosomes) increased conduction velocity and maximum capture rate, along with decreased excitation threshold and improved electrophysiological and Ca 2+ handling properties as shown by response to epinephrine [176]. In another study, a bioreactor was designed to deliver either monophasic (5 V for 2 ms, 1 Hz) or biphasic (+2.5 V for 1 ms, −2.5 V for 1 ms, 1 Hz) electrical pulses with tunable amplitude, pulse width, and frequency to human cardiac progenitors cultured on gelatin coated glass slides [344,361]. After 72 hours, biphasic electrical stimulation resulted in upregulation of early cardiac transcription factors (MEF2D, GATA-4, Nkx2.5), greater expression of late cardiac sarcomeric proteins (cTnT, cardiac α-actinin, and SERCA2a), and earlier expression of Cx43 compared to monophasic electrical stimulation [344].…”

Specific Intensity Direct Current (DC) Electric Field Improves Neural Stem Cell Migration and Enhances Differentiation towards βIII-Tubulin+ Neurons

“…In another example, monophasic and biphasic pulsed electric fields were applied to the human cardiac progenitor cells (hCPCs) isolated from human heart fragment, and induced early differentiation towards a cardiac phenotype. Interestingly, only the biphasic fields showed effectiveness in the up-regulation of cardiac transcription factors [23]. Within the same AC electric field, cell differentiation could be a function of the field frequency.…”