Reconstruction of Endometrium from Human Endometrial Side Population Cell Lines

Irene Cervelló, Aymara Mas, Claudia Gil-Sanchis, Claudia Gil-Sanchís, Laura Peris, Amparo Faus, Philippa T K Saunders, Hilary O D Critchley, Hilary Critchley, Carlos Simón
PloS one · 2011 · vol. 6(6) , pp. e21221 · doi:10.1371/journal.pone.0021221 · PMID:21712999 · PMC3119688 · W2064441872
article OA: gold CC0 ⤵ 56 in-corpus citations
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⚙ AI-generated summary by claude@2026-06, 2026-06-08 ⓘ

Seven human endometrial side population cell lines were established and characterized, demonstrating stem cell features, mesenchymal attributes, and the ability to generate functional human endometrium in vivo.

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⚙ AI-generated deep summary by claude@2026-06, 2026-06-09 ⓘ

This study aimed to establish and characterize human endometrial side population (hESP) somatic stem cell lines derived from the epithelial and stromal compartments using Hoechst 33342 dye efflux, then assess their molecular phenotype and differentiation and regenerative capacity in vivo. Seven hESP cell lines (four epithelial and three stromal) were generated from hypoxia-cloned SP cells, shown to have a normal 46XX karyotype, intermediate telomerase activity, expression of undifferentiated and mesoderm-associated transcripts (including OCT4, NANOG, and WT1), and immunophenotypes consistent with either epithelial (CD9+) or stromal/mesenchymal (vimentin+, CD90+, CD73+, CD45−) origins; they were ERα- and PR-negative and differentiated in vitro into adipocytes and osteocytes. The authors also reported that the hESP lines could reconstruct human endometrium after transplantation beneath the renal capsule of NOD-SCID mice, while noting prior endometrial SP reconstruction efficiencies were low in earlier in vivo experiments. This paper is centrally about endometriosis-independent endometrial stem cell modeling—specifically, it relates to endometriosis by proposing its cell-line system as a model to investigate targets for endometrium proliferation in endometriosis.

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Abstract

Endometrial regeneration is mediated, at least in part, by the existence of a specialized somatic stem cell (SSC) population recently identified by several groups using the side population (SP) technique. We previously demonstrated that endometrial SP displays genotypic, phenotypic and the functional capability to develop human endometrium after subcutaneous injection in NOD-SCID mice. We have now established seven human endometrial SP (hESP) cell lines (ICE 1-7): four from the epithelial and three from the stromal fraction, respectively. SP cell lines were generated under hypoxic conditions based on their cloning efficiency ability, cultured for 12-15 passages (20 weeks) and cryopreserved. Cell lines displayed normal 46XX karyotype, intermediate telomerase activity pattern and expressed mRNAs encoding proteins that are considered characteristic of undifferentiated cells (Oct-4, GDF3, DNMT3B, Nanog, GABR3) and those of mesodermal origin (WT1, Cardiac Actin, Enolase, Globin, REN). Phenotype analysis corroborated their epithelial (CD9+) or stromal (vimentin+) cell origin and mesenchymal (CD90+, CD73+ and CD45⁻) attributes. Markers considered characteristic of ectoderm or endoderm were not detected. Cells did not express either estrogen receptor alpha (ERα) or progesterone receptor (PR). The hESP cell lines were able to differentiate in vitro into adipocytes and osteocytes, which confirmed their mesenchymal origin. Finally, we demonstrated their ability to generate human endometrium when transplanted beneath the renal capsule of NOD-SCID mice. These findings confirm that SP cells exhibit key features of human endometrial SSC and open up new possibilities for the understanding of gynecological disorders such as endometriosis or Asherman syndrome. Our cell lines can be a valuable model to investigate new targets for endometrium proliferation in endometriosis.

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Condition tags

endometriosis

MeSH descriptors

Endometrium Endometrium Regeneration Side-Population Cells Animals Biomarkers Biomarkers Cell Differentiation Cell Line Cell Separation Endometrium Epithelial Cells Epithelial Cells Epithelial Cells Estrogen Receptor alpha Estrogen Receptor alpha Female Humans Karyotyping Mice

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