Int J Stem Cells.  2019 Nov;12(3):440-448. 10.15283/ijsc18139.

Immunosuppressive Effect of Exosomes from Mesenchymal Stromal Cells in Defined Medium on Experimental Colitis

• Ma ZJ
• Wang YH
• Li ZG
• Wang Y
• Li BY
• Kang HY
• Wu XY

• KMID: 2465884
• DOI: http://doi.org/10.15283/ijsc18139

Abstract

BACKGROUND AND OBJECTIVES
The exosomes released by mesenchymal stromal cells (MSCs) in classical FBS-containing media have been demonstrated as an alternative, cell-free therapy in various diseases including inflammatory bowel disease (IBD). It has been found that the function of exosomes is affected by culture condition. We previously developed a serum-free, xeno-free and chemically defined medium, and umbilical cord-derived MSCs in this medium retained the immunosuppressive capability.
METHODS
In this study, we evaluated the immunosuppressive function of exosomes from MSCs (MSC-Exo) in defined medium and their therapeutic effect on treating colitis.
RESULTS
AND CONCLUSIONS: In vitro studies indicated that MSC-Exo reduced the concentration of pro-inflammatory cytokines IFN-γ, TNF-α and IL-1β, and increased the secretion of anti-inflammatory cytokines TGF-β1 and IL-10, but no significant change of inhibitory effect on peripheral blood mononuclear cells proliferation was shown. In vivo experimental colitis showed that administration of MSC-Exo was able to significantly ameliorate the disease activity index score, weight loss, colon shortening, and the histological colitis score through up-regulation anti-inflammatory responses and down-regulation of inflammatory responses. Moreover, the use of MSC-Exo (200 μg) led to an improved therapeutic efficacy when compared with MSCs at a dose of 1×10⁶ cells. Our findings indicate that the exosomes from MSCs in defined medium possess a certain degree of immunosuppressive effect in vitro and exhibit a therapeutic capability in a mouse model of DSS-induced colitis through suppressing inflammation mechanism.

Keyword

Mesenchymal stromal cells; Colitis; Immunosuppressive effect; Serum free; Exosome

MeSH Terms

Animals
Colitis*
Colon
Cytokines
Down-Regulation
Exosomes*
In Vitro Techniques
Inflammation
Inflammatory Bowel Diseases
Interleukin-10
Mesenchymal Stromal Cells*
Mice
Up-Regulation
Weight Loss
Cytokines
Interleukin-10

Figure

• 

  Fig. 1 Characteristics of MSC-Exo. (A) Transmission electron microscopy analysis of exosomes secreted by UCMSCs cultured in defined medium. Scale bar: 50 nm. (B) CD63, CD9, and CD81 expressions in MSC-Exo were detected by western blotting. (C) The size distribution profile of MSC-Exo.

• 

  Fig. 2 MSC-Exo possessed a certain degree of immunosuppressive capability in vitro. The concentrations of the pro-inflammatory cytokines (A) IFN-γ, (B) TNF-α and (C) IL-1β and anti-inflammatory cytokines (D) TGF-β1 and (E) IL-10 were measured in the supernatant of PBMCs treated with different levels of MSC-Exo for 72 h. (F) The proliferation of PBMCs was evaluated after culture with different levels of MSC-Exo. Bars indicate means±SD. n=5; *p<0.05, **p<0.01, and ***p<0.001.

• 

  Fig. 3 MSC-Exo or MSCs alleviated clinical symptom in DSS-induced colitis mice. (A) Scheme of study design. (B) Body weight was monitored daily. (C) Disease activity index (DAI) scores were measured on day 10. Bars indicate means±SD. n=5~8; *p<0.05 and **p<0.01.

• 

  Fig. 4 MSC-Exo or MSCs alleviated colonic damage in DSS-induced colitis mice. (A) Representative images of colon length. (B) Colon length was quantitatively analyzed. (C) Representative H&E staining, bar=100 μm. (D) Corresponding severity score was determined. Bars indicate means± SD. n=5~8; *p<0.05.

• 

  Fig. 5 MSC-Exo or MSCs reduced the infammatory state in DSS-induced colitis mice. The concentrations of the pro-inflammatory cytokines (A) IFN-γ, (B) TNF-α, (C) IL-6 and (D) IL-17 and anti-inflammatory cytokines (E) IL-10 and (F) TGF-β1 in colonic protein extracts were measured by ELISA. Bars indicate mean±SD, n=5~8; *p<0.05, **p<0.01 and ***p<0.001.

Reference

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