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Updated: Dec 29, 2025
Large-Scale Preparation of Synovial Fluid Mesenchymal Stem Cell-Derived Exosomes by 3D Bioreactor Culture
Published on: July 26, 2022
[Experimental study of human amniotic mesenchymal stem cell exosome promoting fibroblasts migration through
Tao Chen1, Shaoying Gao1, Yi Hao1
1Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi Guizhou, 563000, P.R.China.
Objective:
To investigate the effect of microRNA-135a (miR-135a) in human amnion mesenchymal stem cell exosome (hAMSC-Exo) on the migration of fibroblasts.
Methods:
The hAMSC-Exo was extracted with exosomes separation kit and identified, the effect of hAMSC-Exo on fibroblasts migration was detected by scratch test. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the relative expression of miR-135a gene in hAMSC-Exo after overexpression of miR-135a. Scratch test was used to detect the effect of hAMSC-Exo on the migration of fibroblasts after overexpression and knockdown of miR-135a. Western blot was used to detect the migration related proteins of fibroblasts [large tumor suppressor 2 (LATS2), E-cadherin, N-cadherin, and α smooth muscle actin (α-SMA)] after overexpression and knockdown of miR-135a. The 293T cell exosomes and hAMSC-Exo were used as control.
Results:
hAMSC-Exos were extracted successfully. Scratch test results showed that hAMSC group had the strongest ability to promote fibroblasts migration, and GW4869 (exosome inhibitor) treatment group had reduced ability to promote fibroblasts migration. qRT-PCR test showed that the relative expression of miR-135a gene in hAMSC-Exo increased significantly after over expression of miR-135a. Scratch test results showed that after over expression of miR-135a, hAMSC-Exo enhanced the migration ability of fibroblasts, while after knockdown of miR-135a, hAMSC-Exo weakened the migration ability of fibroblasts. Western blot results showed that the expressions of E-cadherin, N-cadherin, LATS2 were down regulated and α-SMA was up regulated in each hAMSC-Exo treatment group when compared with 293T cell exosomes group; after over expression of miR-135a, hAMSC-Exo decreased the expressions of E-cadherin, N-cadherin, LATS2 and increased the expression of α-SMA; while after knockdown of miR-135a, the ability of hAMSC-Exo was weakened.
Conclusion:
miR-135a in hAMSC-Exo can promote fibroblasts' migration, inhibit the expressions of E-cadherin, N-cadherin, LATS2, and promote the expression of α-SMA.
Insights
MicroRNA-135a (miR-135a) within human amnion mesenchymal stem cell exosomes (hAMSC-Exo) promotes fibroblast migration. This involves regulating key proteins like E-cadherin and α-SMA, impacting cellular movement.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Mesenchymal stem cell-derived exosomes are investigated for therapeutic potential.
- Exosomes contain microRNAs that can modulate recipient cell functions.
- Fibroblast migration is crucial in wound healing and tissue repair.
Purpose of the Study:
- To determine the role of microRNA-135a (miR-135a) carried by human amnion mesenchymal stem cell exosomes (hAMSC-Exo) in regulating fibroblast migration.
- To elucidate the molecular mechanisms underlying hAMSC-Exo mediated fibroblast migration.
Main Methods:
- hAMSC-Exosomes were isolated and characterized.
- Fibroblast migration was assessed using scratch assays.
- miR-135a expression levels were manipulated (overexpression and knockdown) in hAMSC-Exo.
- Protein expression of migration-related markers (LATS2, E-cadherin, N-cadherin, α-SMA) was analyzed via Western blot.
Main Results:
- hAMSC-Exo significantly enhanced fibroblast migration.
- Overexpression of miR-135a in hAMSC-Exo further boosted fibroblast migration, while knockdown diminished this effect.
- hAMSC-Exo treatment altered the expression of key proteins: decreased E-cadherin, N-cadherin, and LATS2, and increased α-SMA, with miR-135a levels correlating with these changes.
Conclusions:
- miR-135a within hAMSC-Exo acts as a key mediator promoting fibroblast migration.
- The study demonstrates that miR-135a in hAMSC-Exo influences fibroblast migration by modulating the expression of E-cadherin, N-cadherin, LATS2, and α-SMA.
- These findings highlight the potential of hAMSC-Exo as a therapeutic agent for conditions involving fibroblast migration.
