Anat Cell Biol.  2022 Mar;55(1):100-108. 10.5115/acb.21.169.

Evaluating the effect of conditioned medium from endometrial stem cells on endometriosis-derived endometrial stem cells

Affiliations
  • 1Department of Mesenchymal Stem Cells, The Academic Centre for Education, Culture and Research, Qom, Iran
  • 2Department of Reproductive Biology, The Academic Centre for Education, Culture and Research, Qom, Iran

Abstract

Endometriosis is a common, benign gynecological disease which is determined as an overspreading of endometrial tissue in exterior region of the uterine cavity. Evidence suggests that retrograde menstrual blood which contains mesenchymal stem cells with differential gene expression compared to healthy women may play a role in endometriosis creation. We aimed to identify whether the conditioned medium (CM) from menstrual blood-derived mesenchymal stem cells (MenSCs) of healthy women can affect the expression level of inf lammatory and stemness genes of MenSCs from endometriosis women. Endometriosis-derived MenSCs (E-MenSCs) were treated with CM derived from healthy women’s MenSCs (non-endometriosis derived MenSCs [NE-MenSCs]). Some CD markers were analyzed by flow cytometer before and after treatment compared with NE-MenSCs, and the expression level of inflammatory and stemness genes was evaluated by real-time PCR. E-MenSCs show different morphology in vitro culture in comparison with NE-MenSCs, which were changed in the presence of CM, into a morphology more similar to normal cells and showed significant decrease expression of CD10 after CM treatment. In our results, the interleukin-1, cyclooxygenase-2, and hypoxia-inducible factor 1α as inflamaturay genes and octamer-binding transcription factor 4, NANOG, and sex determining region Y-box 2 as stemness genes showed significantly different expression level in E-MenSCs after treating with CM. Our study indicates that the expression level of some inflammatory- and stemness-related genes which have differential expression in E-MenSCs compared with NEMenSCs, could be changed to normal status by using CM derived from NE-MenSCs.

Keyword

Endometriosis; Mesenchymal stem cells; Conditioned Medium

Figure

  • Fig. 1 The flowchart of the study design. NE-MenSCs, non-endometriosis derived menstrual blood-derived mesenchymal stem cells; E-MenSCs, endometriosis derived menstrual blood-derived mesenchymal stem cells; CM, conditioned medium.

  • Fig. 2 Morphological difference between non-endometriosis derived menstrual blood-derived mesenchymal stem cells (NE-MenSCs) and endometriosis derived menstrual blood-derived mesenchymal stem cells (E-MenSCs). NE-MenSCs in 2D culture showed a fibroblast-like spindle-shaped morphology (A), while E-MenSCs were less stretched and more circular (B). After conditioned medium (CM) treatment, the cells changed to spindle-shaped morphology (C).

  • Fig. 3 (A) The expression of menstrual blood-derived mesenchymal stem cells (MenSCs) surface markers was detected by flow cytometry so that they were positive for CD90, CD10, CD44, CD29, CD73, and CD105, while negative for CD34, CD133, CD45, and CD38. (B) MFI was used to compare E-MenSCs with NE-MenSCs and E-MenSCs after conditioned medium treatment. MFI, mean fluorescence intensity; E-MenSCs, endometriosis derived MenSCs; NE-MenSCs, non endometriosis-derived MenSCs.

  • Fig. 4 Comparison of the inflammatory genes expression in NE-MenSCs, E-MenSCs (before), and E-MenSCs after conditioned medium (CM) treatment (after) by real-time PCR analysis. a)Significant difference between NE-MenSCs versus E-MenSCs. b)Significant difference between before versus after CM treatment. NE-MenSCs, non-endometriosis derived menstrual blood-derived mesenchymal stem cells; E-MenSCs, endometriosis derived menstrual blood-derived mesenchymal stem cells.

  • Fig. 5 Comparison of stemness genes expression in NE-MenSCs, E-MenSCs (before), and E-MenSCs after conditioned medium (CM) treatment (after) by real-time PCR analysis. a)Significant difference between NE-MenSCs versus E-MenSCs. b)Significant difference between before versus after CM treatment. NE-MenSCs, non-endometriosis derived menstrual blood-derived mesenchymal stem cells; E-MenSCs, endometriosis derived menstrual blood-derived mesenchymal stem cells.


Reference

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