Generation of immortalized human endometrial stromal cell lines with different endometriosis risk genotypes

S J Holdsworth-Carson, Sarah J. Holdsworth‐Carson, E M Colgrave, Eliza M Colgrave, J F Donoghue, Jacqueline F. Donoghue, J N Fung, Jenny N. Fung, Molly Churchill, M L Churchill, S Mortlock, Sally Mortlock, P Paiva, Premila Paiva, M Healey, Martin Healey, Grant W. Montgomery, G W Montgomery, J E Girling, Jane E. Girling, P A W Rogers, Peter A. W. Rogers
Molecular human reproduction · 2019 · vol. 25(4) , pp. 194–205 · doi:10.1093/molehr/gaz006 · PMID:30770928 · W2913866795
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Researchers generated immortalized human endometrial stromal cell lines from patients with specific endometriosis risk genotypes, confirming their fibroblast morphology, genotype, and diverse responses to hormones and inflammation.

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Abstract

Endometriotic lesions are composed in part of endometrial-like stromal cells, however, there is a shortage of immortalized human endometrial stromal cultures available for research. As genetic factors play a role in endometriosis risk, it is important that genotype is also incorporated into analysis of pathological mechanisms. Human telomerase reverse transcriptase (hTERT) immortalization (using Lenti-hTERT-green fluorescent protein virus) took place following genotype selection; 13 patients homozygous for either the risk or non-risk 'other' allele for one or more important endometriosis risk single nucleotide polymorphism on chromosome 1p36.12 (rs3820282, rs56318008, rs55938609, rs12037376, rs7521902 or rs12061255). Short tandem repeat DNA profiling validated that donor tissue matched that of the immortalized cell lines and confirmed that cultures were genetically novel. Expression of morphological markers (vimentin and cytokeratin) and key genes of interest (telomerase, estrogen and progesterone receptors and LINC00339) were examined and functional assays for cell proliferation, steroid hormone and inflammatory responses were performed for 7/13 cultures. All endometrial stromal cell lines maintained their fibroblast-like morphology (vimentin-positive) and homozygous endometriosis-risk genotype following introduction of hTERT. Furthermore, the new stromal cultures demonstrated positive and diverse responses to hormones (proliferation and decidualisation changes) and inflammation (dose-dependent response), while maintaining hormone receptor expression. In conclusion, we successfully developed a range of human endometrial stromal cell lines that carry important endometriosis-risk alleles. The wider implications of this approach go beyond advancing endometriosis research; these cell lines will be valuable tools for multiple endometrial pathologies offering a level of genetic and phenotypic diversity not previously available.

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Condition tags

mesh:D004715endometriosis

MeSH descriptors

Endometriosis Founder Effect Genotype Stromal Cells Telomerase Adult Biomarkers Biomarkers Cell Line, Transformed Cell Proliferation Chromosomes, Human, Pair 1 Chromosomes, Human, Pair 1 Chromosomes, Human, Pair 1 Endometriosis Endometriosis Endometriosis Endometrium Endometrium Endometrium Female

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