In vitro exploration of the therapeutic potential of mesenchymal stem cell conditioned medium in endometriosis treatment by targeting apoptosis and cellular migration

Hassan Hedayatnia, Hedayatnia H, Kalhor N, Naser Kalhor
International journal of reproductive biomedicine · 2026 · vol. 24(2) , pp. 117–126 · doi:10.18502/ijrm.v24i2.21052 · PMID:42051963 · PMC13111932
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⚙ AI-generated summary by gemini-2.5-flash-lite, 2026-06-08 ⓘ

Mesenchymal stem cell conditioned media from healthy donors increased apoptosis and inhibited endometriotic cell migration in vitro.

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⚙ AI-generated deep summary by claude@2026-06, 2026-06-10 · read from full text ⓘ

This in vitro experimental study prepared conditioned media (CM) from healthy menstrual blood-derived mesenchymal stem cells (MenSCs) and adipose tissue-derived mesenchymal stem cells (ADSCs), then exposed MenSCs isolated from women with stage III–IV deep endometriosis to these cell-free media. MenSC identity was confirmed by flow cytometry for MSC markers (CD29, CD44, CD73, CD105) and lack of hematopoietic markers, and CM effects were assessed using a scratch wound-closure migration assay and FITC Annexin V/propidium iodide apoptosis flow cytometry, along with RT-qPCR for apoptotic regulators (notably Bax and Bax/Bcl-2). Both ADSC-CM and healthy MenSC-CM significantly reduced scratch area closure (inhibited migration) and increased early and late apoptosis compared with standard DMEM-LG + FBS, accompanied by increased Bax expression and Bax/Bcl-2 ratio. A major limitation is the small donor numbers used to generate CM (healthy donors n=3 and endometriosis donors n=3) and the study’s confinement to in vitro assays. This paper is centrally about endometriosis—specifically, it tests how mesenchymal stem cell conditioned media affects apoptosis and migration in MenSCs derived from women with endometriosis.

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Abstract

BACKGROUND: Endometriosis, a common gynecological disorder involving ectopic endometrial tissue, leads to infertility and chronic pain. Dysregulated apoptosis and abnormal cell migration are key pathological features. Given current treatment limitations, novel strategies like mesenchymal stem cells (MSC) derived conditioned media (CM) are explored due to their rich secretome. OBJECTIVE: To investigate the effects of CM from healthy menstrual blood-derived MSCs (MenSCs-CM) and human adipose tissue-derived MSCs (ADSCs-CM) on apoptosis and migration in endometriotic MenSCs. MATERIALS AND METHODS: This in vitro study involved the isolation of endometriotic MenSCs from infertile women (25-35 yr) via menstrual blood. Mononuclear cells were cultured to passage 3, and characterized using flow cytometry (CD29, CD44, CD73, CD105 positive; CD34, CD38, CD45 negative). CM was prepared separately from healthy donor MenSCs and ADSCs. Endometriotic MenSCs were treated with healthy MenSCs-CM and ADSCs-CM independently. Cell migration (scratch assay), apoptosis (Annexin V), and Bax mRNA levels and Bax/Bcl-2 ratio (real-time polymerase chain reaction) were evaluated. RESULTS: Both ADSCs-CM and MenSCs-CM significantly increased during early and late apoptosis in endometriotic MenSCs (p < 0.001). Scratch assays showed significantly decreased MenSCs migration at 24, 48, and 72 hr (p < 0.001, p = 0.003, p < 0.001, respectively). Gene expression revealed significant increases in Bax mRNA (p = 0.013) and the Bax/Bcl-2 ratio (p = 0.044). CONCLUSION: CM from ADSCs and MenSCs of healthy women enhances apoptosis and inhibits endometriotic MenSCs migration in vitro, suggesting potential therapeutic strategies for endometriosis.
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Both authors designed the study and conducted the research. Also monitored, evaluated, and analyzed the results of the study. Further, they reviewed the article. Both authors approved the final manuscript and take responsibility for the integrity of the data.

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The authors declare that they have no conflict of interest.

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endometriosisinfertility

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