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Abstract Extracellular matrix (ECM) in the human tissue contains vesicles, which are defined as matrix‐bound nanovesicles (MBVs). MBVs serve as one of the functional components in ECM, recapitulating part of the regulatory roles and in vivo microenvironment. In this study, extracellular vesicles from culture supernatants (SuEVs) and MBVs are isolated from the conditioned medium or ECM, respectively, of 3D human mesenchymal stem cells. Nanoparticle tracking analysis shows that MBVs are smaller than SuEVs (100–150 nm). Transmission electron microscopy captures the typical cup shape morphology for both SuEVs and MBVs. Western blot reveals that MBVs have low detection of some SuEV markers such as syntenin‐1. miRNA analysis of MBVs shows that 3D microenvironment enhances the expression of miRNAs such as miR‐19a and miR‐21. In vitro functional analysis shows that MBVs can facilitate human pluripotent stem cell‐derived forebrain organoid recovery after starvation and promote high passage fibroblast proliferation. In macrophage polarization, 2D MBVs tend to suppress the pro‐inflammatory cytokine IL‐12 β , while 3D MBVs tend to enhance the anti‐inflammatory cytokine IL‐10. This study has the significance in advancing the understanding of the bio‐interface of nanovesicles with human tissue and the design of cell‐free therapy for treating neurological disorders such as ischemic stroke.
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Live Mapping of the Brain Extracellular Matrix and Remodeling in Neurological Disorders
Abstract Live imaging of the brain extracellular matrix (ECM) provides vital insights into changes that occur in neurological disorders. Current techniques such as second or third‐harmonic generation offer limited contrast for live imaging of the brain ECM. Here, a new method, pan‐ECM via chemical labeling of extracellular proteins, is introduced for live brain ECM imaging. pan‐ECM labels all major ECM components in live tissue including the interstitial matrix, basement membrane, and perineuronal nets. pan‐ECM enables in vivo observation of the ECM heterogeneity between the glioma core and margin, as well as the assessment of ECM deterioration under stroke condition, without ECM shrinkage from tissue fixation. These findings indicate that the pan‐ECM approach is a novel way to image the entire brain ECM in live brain tissue with optical resolution. pan‐ECM has the potential to advance the understanding of ECM in brain function and neurological diseases.
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- Award ID(s):
- 2123971
- PAR ID:
- 10472593
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Small Methods
- Volume:
- 8
- Issue:
- 1
- ISSN:
- 2366-9608
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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