Adipose Stem Cell-Derived Extracellular Vesicles Induce Proliferation of Schwann Cells via Internalization

Haertinger, Maximilian; Weiss, Tamara; Mann, Anda; Tabi, Annette; Brandel, Victoria; Radtke, Christine

doi:10.3390/cells9010163

Cited by 37 publications

(36 citation statements)

References 61 publications

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“…By the fluorescent intraluminal tagging of EVs with palmNG, we obtained a reliable tool that allowed us to follow their uptake by recipient cells. We have shown that the internalization of melanoma-derived EVs by Huh-7 cells is a time- and dose-dependent process, during which the vesicles localized predominantly to the perinuclear area, as was consistent with other studies [ 44 , 45 , 46 , 47 ]. Based on the analysis of the green fluorescence signal size and distribution, we might speculate that palmNG EVs were trapped in vesicles such as endosomes, transported actively to the perinuclear region, and accumulated in larger organelles.…”

Section: Discussionsupporting

confidence: 93%

Palmitoylated mNeonGreen Protein as a Tool for Visualization and Uptake Studies of Extracellular Vesicles

et al. 2020

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Extracellular vesicles (EVs) are membranous nanoparticles released by cells as vital mediators of intercellular communication. As such, EVs have become an attractive target for pathogens and cancer cells, which can take control over their cargo composition, as well as their trafficking, shaping the pathogenesis. Despite almost four decades of research on EVs, the number of specific and efficient EV labeling methods is limited, and there is still no universal method for the visualization of their transport in living cells. Lipophilic dyes that non-specifically intercalate into the EVs membranes may diffuse to other membranes, leading to the misinterpretation of the results. Here, we propose a palmitoylated fluorescent mNeonGreen (palmNG) protein as an alternative to chemical dyes for EVs visualization. The Branchiostoma lanceolatum-derived mNeonGreen is a brighter, more stable, and less sensitive to laser-induced bleaching alternative to green fluorescent protein (GFP), which makes it a more potent tag in a variety of fluorescence-based techniques. A palmNG-expressing stable human melanoma cell line was generated using retrovirus gene transfer and cell sorting. This protein partially localizes to cellular membranes, and can be detected inside size-exclusion (SEC)-purified EVs. With the use of flow cytometry and fluorescent confocal microscopy, we performed qualitative and quantitative analyses of palmNG-EVs uptake in recipient human hepatoma cells, in comparison to PKH67-labeled vesicles. Our findings confirm that membrane-embedded mNeonGreen can be successfully applied as a tool in EVs transfer and uptake studies.

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Section: Discussionsupporting

confidence: 93%

Palmitoylated mNeonGreen Protein as a Tool for Visualization and Uptake Studies of Extracellular Vesicles

et al. 2020

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“…This procedure enabled to achieve highly enriched rSC ( Figure 1C) and rFB ( Figure 1D) cultures of >95% purity. 75,76 To receive primary cultures of sensory neurons, excised rDRGs were digested and the cell suspension ( Figure 1E) was grown on PDL/laminin coated dishes. The p0 rDRG cultures consisted of rDRG neurons ( Figure 1F, arrowhead), rSCs ( Figure 1F, arrow) as well as rFBs and neurite outgrowth was observed 1 day after seeding.…”

Section: Establishment and Characterization Of Rsc Rfb And Rdrg Nmentioning

confidence: 99%

Defining the regenerative effects of native spider silk fibers on primary Schwann cells, sensory neurons, and nerve‐associated fibroblasts

et al. 2020

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“…The results indicate a comparable regeneration in the allograft group compared to the current clinical gold standard. Furthermore, promising materials such as spider silk [ 81 ], and multipotent cells such as mesenchymal stem cells [ 82 , 83 ], or stem cell-derived extracellular vesicles [ 84 , 85 , 86 ] could further optimize the outcome following allograft nerve repair.…”

Section: Discussionmentioning

confidence: 99%

Reconstruction of Critical Nerve Defects Using Allogenic Nerve Tissue: A Review of Current Approaches

Kornfeld

Borger

Radtke

2021

IJMS

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Regardless of the nerve defect length, nerve injury is a debilitating condition for the affected patient that results in loss of sensory and motor function. These functional impairments can have a profound impact on the patient’s quality of life. Surgical approaches for the treatment of short segment nerve defects are well-established. Autologous nerve transplantation, considered the gold standard, and the use of artificial nerve grafts are safe and successful procedures for short segment nerve defect reconstruction. Long segment nerve defects which extend 3.0 cm or more are more problematic for repair. Methods for reconstruction of long defects are limited. Artificial nerve grafts often fail to regenerate and autologous nerve grafts are limited in length and number. Cadaveric processed/unprocessed nerve allografts are a promising alternative in nerve surgery. This review gives a systematic overview on pre-clinical and clinical approaches in nerve allograft transplantation.

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Adipose Stem Cell-Derived Extracellular Vesicles Induce Proliferation of Schwann Cells via Internalization

Cited by 37 publications

References 61 publications

Palmitoylated mNeonGreen Protein as a Tool for Visualization and Uptake Studies of Extracellular Vesicles

Palmitoylated mNeonGreen Protein as a Tool for Visualization and Uptake Studies of Extracellular Vesicles

Defining the regenerative effects of native spider silk fibers on primary Schwann cells, sensory neurons, and nerve‐associated fibroblasts

Reconstruction of Critical Nerve Defects Using Allogenic Nerve Tissue: A Review of Current Approaches

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