Int J Stem Cells.  2020 Nov;13(3):414-423. 10.15283/ijsc20049.

miR-29a in Exosomes from Bone Marrow Mesenchymal Stem Cells Inhibit Fibrosis during Endometrial Repair of Intrauterine Adhesion

• Tan Q
• Xia D
• Ying X

• KMID: 2508915
• DOI: http://doi.org/10.15283/ijsc20049

Abstract

Background and Objectives
Bone marrow mesenchymal stem cells (BMSCs) is an ideal source of stem cells in the treatment of intrauterine adhesion. Exosomes are a type of membrane vesicle and the diameter is 30∼100 nm. Exosomes can take their contents into the target cells, releasing and exerting their functions. In this study, we intend to study the role of human BMSC-derived exosomes (BMSC-Exo) in promoting endometrial damage repair in the treatment of IUA.
Methods
We used the magnetic bead affinity method to extract BMSC-Exo and analyzed its biological character. Then we co-cultured the BMSCs-Exo with endometrial cells to detect its effect. We injected BMSCs-Exo into the IUA mouse model. We over-expressed miR-29a in BMSCs-Exo by transient transfection, then used RT-PCR to analyze the expression of the related genes.
Results
BMSCs-Exo expressed exosome-specific proteins CD9, CD63, and CD81. BMSCs-Exo could bring the contents into the target cells. BMSCs-Exo can promote endometrial repair in vitro or in vivo. BMSCs-Exo overexpressing miR-29a can reduce αSMA, Collagen I, SMAD2, and SMAD3.
Conclusions
In this study, we successfully isolated BMSCs-Exo and proved its character and biological activity. BMSCs-Exo can promote cell proliferation and cell migration in vitro and can repair damaged endometrium in the IUA model. The presence of miR-29a in BMSCs-Exo may be an important factor in its resistance to fibrosis during endometrial repair of IUA. This study provides new ideas for the treatment of patients with IUA and has important clinical research significance.

Keyword

Bone marrow mesenchymal stem cells; Exosomes; miR-29α; Anti-fibrosis

Figure

• 

  Fig. 1 Culture and identification of BMSCs. (A) Cellular morphology of BMSCs, Scale bar: 40 μm. (B) Growth curve of 3rd BMSCs and 5th BMSCs. (C) Expression of surface antigens was carried out by flow cytometry using CD29, CD44, CD73, CD90, CD105, HLA-DR, CD34, and CD45.

• 

  Fig. 2 Identification of BMSCs derived exosomes. (A) Transmission electron microscopy analysis of BMSCs-Exo, Scale bar: 100 μm. (B) Particle size analysis of BMSCs-Exo. (C) Western blot analysis of exosome-specific proteins CD9, CD63, and CD81 in BMSCs-Exo. (D) Immunofluorescent staining analysis to detect labeled exosomes biological activity.

• 

  Fig. 3 BMSCs-Exo promotes endometrial repair in vitro or in vivo. (A) Growth curve of endometrial cells co-cultured with BMSCs-Exo. (B) BMSCs-Exo promoted cell migration by scratch test. (C) HE staining analysis to detect the ability of BMSCs-Exo to repair endometrial damage in vivo, Scale bar: 100 μm. (D) The number of endometrial glands.

• 

  Fig. 4 The nucleic acid component present in BMSCs-Exo can play a role in inhibiting the formation of fibroblasts. (A) Gel electrophoresis of BMSCs-Exo after lysed by proteinase K or RNase A. (B) Silver nitrate staining of BMSCs-Exo after lysed by proteinase K or RNase A. (C) The expression of fibroblast formation related gene (α-SMA, Collagen I) in TGFβ-induced fibroblasts models, which treated by BMSCs-Exo, RNAse-BMSCs-Exo, PROnase-BMSCs-Exo, and PBS.

• 

  Fig. 5 miR-29a-3p in BMSCs-Exo has an anti-fibrotic role during the repair process. (A) miR-29a overexpressed in BMSCs-Exo used transient transfection. (B) RT-PCR analysis of the expression of fibroblast forma-tion related gene, NC: fibroblast-Exo, miR29a: BMSCs-Exo overexpre-ssing miR-29a, *p<0.05.

Reference

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