Primary culture of endometrial mesenchymal stem cells derived from ectopic lesions of patients with adenomyosis

Purpose

This study aimed to establish a protocol for efficiently isolating and expanding adenomyotic lesion-derived endometrial mesenchymal stem cells (A-eMSCs) in vitro.

Methods

Three different methods—namely, the enzymatic method, the explant method, and the enzymatic explant method—were employed to isolate A-eMSCs. The isolation and expansion efficiencies of these three methods were subsequently compared. The enzymatic explant method was then used, and the transforming growth factor beta type I receptor (TGF-βR1) inhibitor A83-01 was added to the culture medium to evaluate its impact on the isolation and expansion efficiencies of A-eMSCs.

Results

The enzymatic explant method resulted in improved morphology, shorter cell confluence time, and greater SUSD2 enrichment in the isolation of primary endometrial cells compared to the other two methods. The proliferation and differentiation potential of A-eMSCs obtained by sorting primary endometrial cells via the enzymatic explant method were significantly higher than those obtained via the other two methods in vitro. Using the enzymatic explant method, culture medium containing A83-01 further reduced the confluence time of the cells and increased A-eMSCs enrichment during the primary endometrial cell isolation stage. Furthermore, A83-01 enhanced the proliferation and maintained the differentiation potential of A-eMSCs during the cell expansion stage.

Conclusion

Our study identified a robust, cost-effective, and efficient protocol for isolating and expanding A-eMSCs and providing an important foundation for further research on the pathogenesis and clinical treatment of AM.