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Related Experiment Video

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Enzymatic Isolation of Skeletal Muscle Interstitial Extracellular Vesicles
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Engineering Extracellular Vesicles to Modulate Their Innate Mitochondrial Load.

Kandarp M Dave1, Duncan X Dobbins1, Maura N Farinelli1,2

  • 1Graduate School of Pharmaceutical Sciences, Duquesne University, 600 Forbes Avenue, 453 Mellon Hall, Pittsburgh, PA USA.

Cellular and Molecular Bioengineering
|November 29, 2022
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Summary
This summary is machine-generated.

Mitochondria-enriched extracellular vesicles (EVs), specifically microvesicles (MVs), can be generated by enhancing mitochondrial biogenesis in donor cells. These enhanced MVs effectively transfer mitochondria to recipient cells, improving their ATP levels and function.

Keywords:
ATPBrain endothelial cellsEVsExosomesMetabolic functionMicrovesiclesMitochondriaPGC-1α

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Neuroscience

Background:

  • Extracellular vesicles (EVs) are investigated as therapeutic delivery vehicles.
  • Mitochondria within EVs can be transferred to recipient cells.
  • Enhancing mitochondrial biogenesis in donor cells may increase mitochondrial load in EVs.

Purpose of the Study:

  • To investigate if increasing mitochondrial biogenesis in donor cells enriches EVs with mitochondria.
  • To determine if mitochondria-enriched EVs (mito-EVs) enhance ATP levels in recipient brain endothelial cells (BECs).

Main Methods:

  • Donor cells (NIH/3T3, hCMEC/D3) were treated with resveratrol to induce mitochondrial biogenesis.
  • EVs (microvesicles and exosomes) were isolated and characterized.
  • Recipient BECs were treated with EVs, and ATP levels were measured.
  • EV uptake and mitochondrial colocalization were assessed using microscopy and flow cytometry.

Main Results:

  • Resveratrol treatment increased mitochondrial markers and mitochondrial content in microvesicles (MVs), but not exosomes.
  • Mito-MVs showed increased particle size and concentration compared to naïve MVs.
  • Mito-EVs successfully transferred mitochondria to recipient BECs, increasing their ATP levels.
  • Mitochondria from mito-MVs colocalized with recipient BEC mitochondria.

Conclusions:

  • Pharmacological modulation of donor cell mitochondrial biogenesis effectively enriches MVs with mitochondria.
  • Mito-MVs demonstrate superior potential for improving mitochondrial function compared to naïve MVs.
  • Mito-EVs hold promise for treating neurovascular and neurodegenerative diseases.