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. 2023 Jun 20;14(1):165.
doi: 10.1186/s13287-023-03397-2.

Comparison of the therapeutic effects between stem cells and exosomes in primary ovarian insufficiency: as promising as cells but different persistency and dosage

Hang-Soo Park  1 , Rishi Man Chugh  2   3 , Jin Seok  1 , Esra Cetin  1 , Hanaa Mohammed  1   4 , Hiba Siblini  1 , Farzana Liakath Ali  1 , Mohammad Mousaei Ghasroldasht  1 , Hiba Alkelani  1 , Amro Elsharoud  2 , Mara Ulin  2 , Sahar Esfandyari  2 , Ayman Al-Hendy  5   6
Affiliations

Comparison of the therapeutic effects between stem cells and exosomes in primary ovarian insufficiency: as promising as cells but different persistency and dosage

Hang-Soo Park et al. Stem Cell Res Ther. .

Abstract

Background: Primary ovarian insufficiency (POI) refers to the loss of ovarian function under the age of 40 and results in amenorrhea and infertility. Our previous studies have shown that transplantation of mesenchymal stem cells (MSCs) and MSC-derived exosomes in chemotherapy-induced POI mouse ovaries can reverse the POI and eventually achieve pregnancy. Based on our recent studies, MSC-derived exosomes have almost equal therapeutic potentials as transplanted MSCs. However, it is still unclear whether exosomes can completely replace MSCs in POI treatment. For the reliable application of cell-free treatment for POI patients using exosomes, there is a need to understand whether there is any outcome and effectiveness difference between MSC and MSC-derived exosome treatment.

Methods: Comparing the therapeutic effect of intravenous injection using MSCs and equal amounts of exosomes in a POI mouse model will reveal the difference between the two therapeutic resources. In this study, we induced POI in C57/BL6 mice by chemotherapy (CXT) using a standard protocol. We then injected four different doses of MSCs or equal amounts of commercialized MSC-derived exosomes by retro-orbital injection post-CXT.

Result: After MSC/exosome treatment, tissue and serum samples were harvested to analyze molecular changes after treatment, while other mice in parallel experiments underwent breeding experiments to compare the restoration of fertility. Both the MSC- and exosome-treated groups had a restored estrous cycle and serum hormone levels compared to untreated POI mice. The pregnancy rate in the MSC-treated group was 60-100% after treatment, while the pregnancy rate in the exosome-treated group was 30-50% after treatment. Interestingly, in terms of long-term effects, MSC-treated mice still showed a 60-80% pregnancy rate in the second round of breeding, while the exosome-treated group became infertile again in the second round of breeding.

Conclusions: Although there were some differences in the efficacy between MSC treatment and exosome treatment, both treatments were able to achieve pregnancy in the POI mouse model. In conclusion, we report that MSC-derived exosomes are a promising therapeutic option to restore ovarian function in POI conditions similar to treatment with MSCs.

Keywords: Exosome; Infertility; Long-term effect; Mesenchymal stem cell; Primary ovarian insufficiency.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Restored estrous cycle in the MSC-injected POI mouse model. Three different numbers of MSCs (MSC-10K: 10,000 cells/100 µL, MSC-100K: 100,000 cells/100 µL, and MSC-1M: 1,000,000 cells/100 µL) were injected intravenously. A Representative image of the proestrus, estrus, metestrus, and diestrus phases in mouse vaginal smear samples. Animal shows significantly delayed cycle (> 95% CI) is highlighted with ǂ symbol. B The estrous cycle length in each mouse after MSC treatment. C Representation of the estrous cycle change in mice by daily vaginal smear analysis
Fig. 2
Fig. 2
Restored ovarian function by MSC treatment. A Representative mouse ovarian tissue image stained with H&E. B Average size of the mouse ovaries in healthy mice (control), untreated POI mice (POI), and MSC-treated POI mice (MSC) (n = 3 per group). C–E Average serum E2, AMH, and FSH levels in the control, POI, and MSC groups (n = 3 per group). F Number of ovarian follicles in the control, POI, and MSC groups. Primordial/primary follicles, secondary follicles, preantral/antral follicles, and corpus lutea were differentially counted and merged into one stacked bar (n = 3 per group)
Fig. 3
Fig. 3
Fertility was restored by MSC treatment in the POI mouse model (n = 6 per group). A Pregnancy rate comparison between healthy mice (control), untreated POI mice (POI), and MSC-treated POI mice (MSC) in the first breeding experiment. B Pregnancy rate comparison between the control group, POI group, and MSC group in the second breeding experiment. C Average number of delivered pups per litter. D Representative image of delivered pups at Day 0. E Genomic DNA PCR analysis for detecting injected cells (human MSCs) in mouse whole blood using human-specific ALU primers (n = 3 per group). F Genomic DNA PCR analysis for detecting injected cells in neonatal offspring liver tissue using human-specific ALU primers (n = 5 per group). Data are presented as the mean ± SD. (Significance level, *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; NS: Not significant.)
Fig. 4
Fig. 4
Effect of MSC-derived exosomes in an in vitro POI model on damaged human granulosa cells (HGrC1). The therapeutic effect in HGrC1 cells was compared between untreated control (control), MSC conditioned media treatment (MSC CM), and MSC-derived exosome treatment (exosomes). A Morphology of HGrC1 cells in the control, MSC CM, and exosome groups after 24 h and 48 h of treatment. B Average number of HGrC1 cells after MSC CM and exosome treatment. C, D Steroid gene expression levels (C: Cyp19, D: StAR) in HGrC1 cells after 24 h of treatment with MSC CM and exosomes. E–K Protein expression levels of the apoptosis marker caspase-3 (E–G) and steroidogenesis markers (I–K) in western blot analysis. Data are presented as the mean ± SD. (n = 3, significance level, *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; NS: Not significant.)
Fig. 5
Fig. 5
Ovarian function restoration in the POI mouse model after exosome treatment. The therapeutic effect in the POI mouse model was compared between the healthy control (control), untreated POI (POI), umbilical-derived MSC exosome treatment (UC-Exo), and bone marrow-derived MSC-derived exosome treatment (BM-Exo) groups. A Estrous cycle after exosome treatment by daily vaginal smear analysis. Animal shows significantly delayed cycle (> 95% CI) is highlighted with ǂ symbol. B Average serum E2, AMH, and FSH levels in the control, POI, UC-Exo, and BM-Exo groups at 2 weeks after treatment. C Representative image of ovarian tissue with H&E staining. D Average size of the ovaries in the control, POI, UC-Exo, and BM-Exo groups at 2 weeks after treatment. E Number of ovarian follicles in the control, POI, UC-Exo, and BM-Exo groups at 2 weeks after treatment. F TUNEL assay in ovarian tissue among the control, POI, UC-Exo, and BM-Exo groups at 2 weeks after treatment. G, H Picrosirius Red staining assay in ovarian tissue among the control, POI, UC-Exo, and BM-Exo groups at 2 weeks after treatment (small image magnification: 50×; large image magnification: 400×). Data are presented as the mean ± SD. (n = 3, significance level, *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; NS: Not significant.)
Fig. 6
Fig. 6
Fertility restoration in the POI mouse model after exosome treatment. A Pregnancy rate in the healthy control (control), untreated POI (POI), umbilical-derived MSC exosome treatment (UC-Exo), and bone marrow-derived MSC-derived exosome treatment (BM-Exo) groups in the first breeding. B Average number of pups per litter between the control, POI, UC-Exo, and BM-Exo groups in the first breeding. C Postnatal growth rate comparison between the control, UC-Exo, and BM-Exo groups. D Representative image of delivered pups on Day 0. E Pregnancy rate of the control, POI, UC-Exo, and BM-Exo groups in the second breeding. F Average number of pups per litter between the control, POI, UC-Exo, and BM-Exo groups in the second breeding. G Ovarian tissue morphology after the second breeding. H–J Serum hormone levels after the second breeding. Data are presented as the mean ± SD. (n = 3, significance level, *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; NS: not significant.)
Fig. 7
Fig. 7
Differences between MSC treatment and exosome treatment in the POI mouse model. A Number of ovarian follicles in the healthy control (control), untreated POI group (POI), MSC treatment group (MSC 1×), equal amount of exosomes (UC-Exo 1×, BM-Exo 1×), tenfold higher amount of exosomes treatment (UC-Exo 10×, BM-Exo 10×), and 100-fold higher amount of exosomes treatment (UC-Exo 100×, BM-Exo 100X). B Comparison of the average number of total follicles among the control, POI, MSC 1×, average UC-Exo and BM-Exo (exosomes 1×, 10×, and 100×) groups. C Altered gene expression by RNA-seq analysis in each treatment compared with untreated POI mouse ovaries. A volcano plot shows significantly increased genes (blue) and significantly decreased genes (red) after MSC and exosome treatment. Data are presented as the mean ± SD. (n = 3, significance level, *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; NS: Not significant.)
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References

    1. Beck-Peccoz P, Persani L. Premature ovarian failure. Orphanet J Rare Dis. 2006;1:9. doi: 10.1186/1750-1172-1-9. - DOI - PMC - PubMed
    1. Christin-Maitre S, Braham R. General mechanisms of premature ovarian failure and clinical check-up. Gynecol Obstet Fertil. 2008;36(9):857–861. doi: 10.1016/j.gyobfe.2008.07.003. - DOI - PubMed
    1. Panay N, Fenton A. Premature ovarian failure: a growing concern. Climacteric. 2008;11(1):1–3. doi: 10.1080/13697130701878635. - DOI - PubMed
    1. Elfayomy AK, Almasry SM, El-Tarhouny SA, Eldomiaty MA. Human umbilical cord blood-mesenchymal stem cells transplantation renovates the ovarian surface epithelium in a rat model of premature ovarian failure: possible direct and indirect effects. Tissue Cell. 2016;48(4):370–382. doi: 10.1016/j.tice.2016.05.001. - DOI - PubMed
    1. Blumenfeld Z, Evron A. Endocrine prevention of chemotherapy-induced ovarian failure. Curr Opin Obstet Gynecol. 2016;28(4):223–229. doi: 10.1097/GCO.0000000000000278. - DOI - PubMed

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