Establishment and characterization of cell lines from human endometrial epithelial and mesenchymal cells from patients with endometriosis

Reproductive Diseases
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VOLUME 1, ISSUE 2, P195-205, NOVEMBER 01, 2020

Authors:

Ayako Muraoka, M.D., Satoko Osuka, M.D., Ph.D., Tohru Kiyono, M.D., Ph.D., Miho Suzuki, Ph.D., Akira Yokoi, M.D., Ph.D., Tomohiko Murase, M.D., Ph.D., Kimihiro Nishino, M.D., Ph.D., Kaoru Niimi, M.D., Ph.D., Tomoko Nakamura, M.D., Ph.D., Maki Goto, M.D., Ph.D., Hiroaki Kajiyama, M.D., Ph.D., Yutaka Kondo, M.D., Ph.D., Fumitaka Kikkawa, M.D., Ph.D.

Abstract:

Objective

To establish and characterize cell lines derived from human endometrial epithelial cells (ECs) and mesenchymal cells (MCs) from patients with and without endometriosis.


Design

In vitro experimental study.

Setting

University and national cancer center research institute.


Patient(s)

Two women with endometriosis and two women without endometriosis.


Intervention(s)

Sampling of endometrial ECs and MCs.


Main Outcome Measure(s)

Establishing immortalized endometrial ECs and MCs with quantitative reverse transcription-polymerase chain reaction (qRT-PCR), immunocytochemical analysis, and RNA sequence profiling performed to characterize the immortalized cells and a cell proliferation assay, three-dimensional culture, and assays for hormone responses performed to characterize the features of ECs.


Result(s)

The qRT-PCR, immunocytochemical analysis, and Western blot analysis revealed that the ECs and MCs maintained their original features. Moreover, the immortalized cells were found to retain responsiveness to sex steroid hormones. The ECs formed a gland-like structure in three-dimensional culture, indicating the maintenance of normal EC phenotypes. The RNA sequence profiling, principal component analysis, and clustering analysis showed that the gene expression patterns of the immortalized cells were different from those of cancer cells. Several signaling pathways that were statistically significantly enriched in ECs and MCs with endometriosis were revealed.


Conclusion(s)

We successfully obtained four paired immortalized endometrial ECs and MCs from patients with and without endometriosis. Using these cells could help identify diagnostic and therapeutic targets for endometriosis. The cell lines established in this study will thus serve as powerful experimental tools in the study of endometriosis.

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