High frequency acoustic cell stimulation promotes exosome generation regulated by a calcium-dependent mechanism

Abstract

Exosomes are promising disease diagnostic markers and drug delivery vehicles, although their use in practice is limited by insufficient homogeneous quantities that can be produced. We reveal that exposing cells to high frequency acoustic irradiation stimulates their generation without detriment to cell viability by exploiting their innate membrane repair mechanism, wherein the enhanced recruitment of calcium ions from the extracellular milieu into the cells triggers an ESCRT pathway known to orchestrate exosomal production. Given the high post-irradiation cell viabilities (≈95%), we are able to recycle the cells through iterative irradiation and post-excitation incubation steps, which facilitate high throughput production of a homogeneous population of exosomes—a particular challenge for translating exosome therapy into clinical practice. In particular, we show that approximately eight- to ten-fold enrichment in the number of exosomes produced can be achieved with just 7 cycles over 280 mins, equivalent to a yield of around 1.7–2.1-fold/h.

Mac-1 Receptor Clustering Initiates Production of Pro-Inflammatory, Antibacterial Extracellular Vesicles From Neutrophils

“…Interestingly, the Ca 2+ ionophore was able to initiate significant EV production also in the absence of external Ca 2+ , probably by mobilization of internal sources. This observation suggests that the machinery itself that leads to physical formation of EVs depends strongly on Ca 2+ signaling independent from the source of Ca 2+ (36)(37)(38).…”

Emerging biogenesis technologies of extracellular vesicles for tissue regenerative therapeutics

“…Other mechanical forces have also been introduced to cells, such as acoustic vibration, to increase the EVs secretion (Figure 4(b)). 48 When a high-frequency acoustic vibration (10 MHz) was applied to cells using an electrical stimulating piezoelectric device, the EVs were produced significantly more (15 times) when compared to non-vibration conditions. The higher EVs production was reasoned to the increased influx of calcium ions.…”

“…applying physicomechanical cues such as forces 35,[46][47][48] and other stimuli (e.g. electricity, thermal, photodynamic, radiative stress).…”

“…The higher EVs production was reasoned to the increased influx of calcium ions. 48 The external mechanical stimuli might alter the cell membrane mechanics and the ion channels, and the intracellular mechanosensitive machineries that are involved in the EVs genesis, 25,[65][66][67] which yet needs future investigation.…”

Alteration of Payload in Extracellular Vesicles by Crosstalk With Mesenchymal Stem Cells From Different Origin

“…Various studies have reported the relief of in ammation in skin tissues and infected organs in animal models [22]. The current focus of antiin ammatory studies using MSC-derived EVs is to increase the payload of useful factors in EVs through discovery of factors through wound healing and mechanism research [23,24]; there are reports that a calcium-dependent mechanism increases the release of EVs from the cells [25]. However, the important aspect of MSC research is to observe the change in payload inside the MSC and understands its role in disease [21].…”