Establishment of an Immortalized Endometriotic Stromal Cell Line from Human Ovarian Endometrioma
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ENDOMETRIOSIS: ORIGINAL ARTICLE
Establishment of an Immortalized Endometriotic Stromal Cell Line from Human Ovarian Endometrioma Yong Song 1 & Niraj R. Joshi 1 & Erin Vegter 1 & Samantha Hrbek 1 & Bruce A. Lessey 2 & Asgerally T. Fazleabas 1 Received: 20 February 2020 / Accepted: 28 May 2020 # Society for Reproductive Investigation 2020
Abstract Endometrial-like stromal cells, one of the main components of endometriotic lesions, are an important in vitro model for studying cellular and molecular mechanisms associated with lesion development in endometriosis. However, the short life span of primary endometriotic stromal cells (Ec-ESCs) limits their use. Human telomerase reverse transcriptase (hTERT) plasmids can be used to develop immortalized cell lines. Here we aimed to establish an endometriotic stromal cell line by hTERT immortalization. Primary Ec-ESCs were obtained from a human ovarian endometriotic cyst. The purity was assessed by morphology and the expression of vimentin, cytokeratin, and human interferon-inducible transmembrane protein 1 (hIFITM1). Cells were infected with hTERT lentiviral vector and selected with hygromycin. hTERT mRNA levels were confirmed by RT-qPCR. Immortalized Ec-ESCs (iEc-ESCs) were characterized by examining the expression of morphological markers and key genes of interest, TP53, estrogen receptor β (ERβ), progesterone receptor (PR), and steroidogenic factor-1 (SF-1). Karyotyping and in vitro decidualization studies were also performed. Ec-ESCs were positive for vimentin and hIFITM1 and negative for cytokeratin, indicating that they were representative of Ec-ESC. The fibroblast-like morphology, expression of TP53, ERβ, PR, and SF-1 did not change before and after hTERT immortalization. iEc-ESCs showed an impaired decidualization response like primary EcESCs when compared to normal eutopic stromal cells. Karyotyping showed that 15/19 cells had normal female karyotype, while 4/19 cells had partial trisomy 11q. Collectively, we successfully established and characterized an immortalized endometriotic stromal cell line. It is potentially useful as an in vitro experimental model to investigate endometriosis biology. Keywords Endometriosis . Endometriotic stromal cells . hTERT . Immortalization . Decidualization
Introduction Endometriosis is a chronic gynecological disorder in women of reproductive age. It is characterized by the presence of endometrial-like tissue outside of the uterine cavity [1]. Endometriosis usually develops in the pelvic and abdominal organs and/cavities, such as the peritoneal wall, ovaries, and fallopian tubes [1]. It is generally categorized into three Electronic supplementary material The online version of this article (https://doi.org/10.1007/s43032-020-00228-0) contains supplementary material, which is available to authorized users. * Asgerally T. Fazleabas [email protected] 1
Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, Grand Rapids, MI 49503, USA
2
Center for Fertility, Endocrinology and Menopause, Wake Forest
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