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. 2024 Jul 26;10(15):e35343.
doi: 10.1016/j.heliyon.2024.e35343. eCollection 2024 Aug 15.

Poly (I:C) increases the expression of galectin 1, 3, 9 and HGF genes in exosomes isolated from human Wharton's jelly mesenchymal stem cells

Mehdi Abbaspour  1 , Mehri Ghafourian Boroujerdnia  1   2 , Mohammad Taher Tahoori  3   4 , Mojtaba Oraki Kohshour  1 , Mohammad Ghasemi Dehcheshmeh  1 , Sareh Amirzadeh  5 , Afshin Amari  1   6
Affiliations

Poly (I:C) increases the expression of galectin 1, 3, 9 and HGF genes in exosomes isolated from human Wharton's jelly mesenchymal stem cells

Mehdi Abbaspour et al. Heliyon. .

Abstract

Background: Mesenchymal stem cells (MSCs) are commonly employed as a powerful tool for the treatment of immune-mediated problems owing to their capacity to regulate the immune system and differentiate into different tissues. Researchers use mesenchymal stem cell products given the limitations associated with the application of MSCs. Exosomes are nanometer vesicles derived from MSCs that are used in cell-free therapy. Inflammatory environmental conditions, such as stimulation of Toll-like receptor 3 (TLR-3), has the ability to adjust the immune-regulating properties and anti-inflammatory function of mesenchymal stem cells and their exosomes. Galectins and hepatocyte growth factor (HGF) are known as immunomodulatory factors in mesenchymal stem cells. This study was designed to examine the expression of galectin-1, galectin-3, galectin-9, and HGF genes in exosomes isolated from human Wharton's jelly mesenchymal stem cells (hWJ-MSCs) after stimulation with Poly (I:C) (Polyinosinic:polycytidylic acid sodium salt).

Methods: To begin, the explant technique was used to extract mesenchymal stem cells from human umbilical cord Wharton's jelly. Then, the stem cells were stimulated using Poly (I:C) at three time intervals of 12, 24 and 48 h. Exosomes secreted from the supernatant of cells were extracted and exosome confirmation tests, including Scanning electron microscopy (SEM), Dynamic light scattering (DLS) and Flow cytometry were performed. Finally, the expression of galectin-1, galectin-3, galectin-9, and HGF genes in exosomes was evaluated by Real-Time PCR at three time intervals of 12, 24 and 48 h after stimulation.

Results: The findings of our study indicated that following stimulation with Poly (I:C), the expression of galectin-9 and HGF (P < 0.05) genes was markedly higher than in the control group after 12 h. After 24 h, the expression of galectin-9 (P < 0.01), galectin-3 and HGF (P < 0.05) increased; the expression of galectin-1, galectin-3, (P < 0.05), galectin-9 and HGF genes (p < 0.01) significantly increased compared to the control group after 48 h.

Conclusion: TLR3 stimulation can increase the expression of galectins and HGF genes in exosomes derived from hWJ-MSCs and may be improve the immunosuppressive abilities of exosomes.

Keywords: Exosome; Galectin; Hepatocyte growth factor; Mesenchymal stem cells; Toll-like receptor 3.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Isolation and characterization of hWJ-MSC. (A) Morphology of WJ-MSCs after about four weeks (B) After 21 days, osteogenic differentiation was assessed by Alizarin Red S staining, calcium deposition was stained bright orange-red. (C) Oil Red O staining of WJ-MSCs, intracellular lipid accumulation was stained bright red in adipocytes at day 21. Original magnifications = 40 × , bar = 200 μm (A–C).
Fig. 2
Fig. 2
Flow cytometry analysis of surface markers of hWJ-MSC. Flow cytometry analysis showing that WJ-MSC were negative for CD34, CD31 and CD45 but positive for CD73, CD90 and CD105. Positive expression was based on isotype control (blue curve).
Fig. 3
Fig. 3
Characterization of exosomes isolated from WJ-MSCs. (A) Size, PDI and zeta potential of exosomes. Data presented as mean ± SD. (B) Dynamic light scattering (DLS) indicated that the mean size of isolated exosomes was 132 nm.
Fig. 4
Fig. 4
Characterization of exosomes by SEM. Scanning electron microscopy (SEM) images indicated that exosomes had a uniform spherical shape with no significant deformities.
Fig. 5
Fig. 5
Flow cytometry analysis of surface markers of exosomes. Flow cytometry analysis showing that exosomes were negative for CD9, CD63 and CD81. Positive expression was based on control (red curve).
Fig. 6
Fig. 6
Toxicity of Poly (I:C) on WJ-MSCs. The viability was greater than 86 % after 12, 24, 48, at different concentration of Poly (I:C). Mean ± SD was used to represent the data.
Fig. 7
Fig. 7
Expression of Galectin-1, Galectin-3, Galectin -9, and HGF genes in exosomes isolated from WJ-MSCs stimulated Poly (I:C). (A) Expression the genes after 12 h. (B) Expression the genes after 24 h. (C) Expression the genes after 48 h*P < 0.05, **P < 0.01. Bars and whiskers represent the mean ± SD. One-way ANOVA was performed to identify the difference between groups.
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