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A method for isolating and culturing ectopic epithelial and stromal cells to study human adenomyosis

  • General Gynecology
  • Published:
Archives of Gynecology and Obstetrics Aims and scope Submit manuscript

Abstract

Purpose

Although adenomyosis is a common and benign gynecological disease, the specific pathogenesis of this condition is yet to be fully elucidated. It is difficult to culture primary cells of the ectopic endometrial epithelia and stroma from human adenomyosis lesions. Most of the previous of studies on adenomyosis were based on primary eutopic endometrium cells. However, as yet, no efficient protocols have been developed for the isolation, culture or purification of primary ectopic epithelial and stromal cells from human adenomyosis lesions. Therefore, the present study aimed to develop an efficient protocol for the isolation and culture of primary ectopic epithelial and stromal cells from human adenomyosis lesions.

Methods

In the present study, we aimed to obtain ectopic endometrium tissue from human adenomyosis foci and use a simple and operable type I collagenase digestion method for primary culture. Cells were isolated by sterile cell strainer filtration and flow cytometry was performed to identify, purify, and evaluate the viability of isolated ectopic endometrial cells.

Results

Using our method, we successfully isolated and cultured highly purified and active ectopic endometrial epithelial and stromal cells from human adenomyosis foci. Ep-CAM was expressed in ectopic epithelial cells of human adenomyosis with a purity of 93.74% and a viability of 80.58%. In addition, CD10 were robustly expressed by ectopic stromal cells in human adenomyosis. Cellular purity and viability were determined to be 96.37 and 93.49%, respectively.

Conclusion

Our method provides a new experimental model for studying the molecular pathogenesis of human adenomyosis.

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Data availability

All the data we did were uploaded and saved in our hospital’s library. As it involves patient privacy and other relevant national policies, it is not provided in this manuscript. However, if anyone is interested in it, please contact the corresponding author directly.

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Acknowledgements

This work was funded by National Key R&D Program of China (Grant number: 2022YFC2704003), National Natural Science Foundation of China (Grant numbers: 81974225, 82001518 and 82171636).

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Authors and Affiliations

  1. Women’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People’s Republic of China

    Zhou Fang, Jianzhang Wang, Tiantian Li, Meichen Yin & Xinmei Zhang

  2. The First Affiliated Hospital of Huzhou University, Huzhou, Zhejiang, People’s Republic of China

    Zhou Fang

  3. Taizhou First People’s Hospital, Huangyan Hospital of Wenzhou Medical University, Taizhou, Zhejiang, People’s Republic of China

    Yangying Peng

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  1. Zhou Fang
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  2. Jianzhang Wang
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  6. Xinmei Zhang
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Contributions

ZF: conceptualization, writing—original draft, data curation, methodology, writing—review and editing, validation, resources. JW and TL: conceptualization, methodology, writing—review and editing, validation, supervision. MY and YP: data curation, methodology, resources, validation. XZ: supervision, conceptualization, writing—review and editing, project administration, validation. All the authors read the submitted version and approved it.

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Correspondence to Xinmei Zhang.

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The authors report no conflict of interest. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Fang, Z., Wang, J., Li, T. et al. A method for isolating and culturing ectopic epithelial and stromal cells to study human adenomyosis. Arch Gynecol Obstet 309, 551–563 (2024). https://doi.org/10.1007/s00404-023-07254-8

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  • DOI: https://doi.org/10.1007/s00404-023-07254-8

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