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Updated: Jul 8, 2025
Isolation, Characterization, and Therapeutic Application of Extracellular Vesicles from Cultured Human Mesenchymal Stem Cells
Published on: September 23, 2022
CD73-positive pediatric urethral mesenchymal stem-like cell-derived small extracellular vesicles stimulate
Shilin Zhang1, Jierong Li1, Chunjing Li1
1Department of Urology, Foshan Maternity & Child Healthcare Hospital, Foshan 528000, China.
Insights
Pediatric urethral stem cell-derived extracellular vesicles promote blood vessel growth by delivering CD73, which activates the adenosine pathway. This finding offers a novel therapeutic strategy for urethral injury repair.
Area of Science:
- Regenerative Medicine
- Vascular Biology
- Biotechnology
Background:
- Angiogenesis is crucial for repairing urethral injuries.
- Stem cells and their secretomes can enhance angiogenesis.
- Pediatric urethral mesenchymal stem-like cells (PU-MSLCs) have been identified.
Purpose of the Study:
- To investigate the pro-angiogenic effects of PU-MSLC-derived small extracellular vesicles (PUMSLC-sEVs).
- To elucidate the underlying mechanisms of PUMSLC-sEVs in promoting angiogenesis.
Main Methods:
- Cultivation and identification of PUMSLCs and PUMSLC-sEVs.
- In vitro assays: EdU, CCK-8, scratch wound, Transwell, and tube formation assays.
- Investigation of the role of CD73 and adenosine (ADO) in PUMSLC-sEV-mediated angiogenesis.
Main Results:
- PUMSLC-sEVs significantly enhanced human umbilical vein endothelial cell (HUVEC) proliferation, migration, and tube formation.
- PUMSLC-sEVs deliver CD73 to HUVECs, promoting angiogenesis.
- Extracellular adenosine (ADO) enhanced HUVEC angiogenic ability, with PUMSLC-sEVs further promoting it via Adenosine Receptor A2A (A2AR) activation.
Conclusions:
- PUMSLC-sEVs possess significant pro-angiogenic properties.
- The mechanism involves the CD73/ADO/A2AR signaling axis.
- PUMSLC-sEVs represent a potential therapeutic agent for promoting angiogenesis in urethral repair.
Introduction:
Angiogenesis plays an important role in the repair of urethral injury, and stem cells and their secretomes can promote angiogenesis. We obtained pediatric urethral mesenchymal stem-like cells (PU-MSLCs) in an earlier study. This project studied the pro-angiogenic effect of PU-MSLC-derived small extracellular vesicles (PUMSLC-sEVs) and the underlying mechanisms.
Materials And Methods:
PUMSLCs and PUMSLC-sEVs were cultivated and identified. Then, biological methods such as the ethynyl deoxyuridine (EdU) incorporation assay, Cell Counting Kit-8 (CCK-8) assay, scratch wound assay, Transwell assay, and tube formation assay were used to study the effect of PUMSLC-sEVs on the proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs). We explored whether the proangiogenic effect of PUMSLC-sEVs is related to CD73 and whether adenosine (ADO, a CD73 metabolite) promoted angiogenesis. GraphPad Prism 8 software was used for data analysis.
Results:
We observed that PUMSLC-sEVs significantly promoted the proliferation, migration, and tube-forming abilities of HUVECs. PUMSLC-sEVs delivered CD73 molecules to HUVECs to promote angiogenesis. The angiogenic ability of HUVECs was enhanced after treatment with extracellular ADO produced by CD73, and PUMSLC-sEVs further promoted angiogenesis by activating Adenosine Receptor A2A (A2AR).
Conclusions:
These observations suggest that PUMSLC-sEVs promote angiogenesis, possibly through activation of the CD73/ADO/A2AR signaling axis.
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