Human endometrial fibroblasts immortalized by simian virus 40 large T antigen differentiate in response to a decidualization stimulus.

Abstract

Human endometrial fibroblasts have been immortalized by infection with simian virus 40 large T antigen and established as a permanent cell line, St-2. Biochemical differentiation of this cell line has been demonstrated by the ability of a decidualizing stimulus, 8-bromo-cAMP plus medroxyprogesterone acetate (MPA), to induce PRL secretion and increase the enzymatic activity of estrone sulfatase. MPA, alone or in combination with estradiol, was unable to elicit this response, but potentiated the effect of 8-bromo-cAMP on PRL production and estrone sulfatase activity. The increase in PRL protein was accompanied by an increase in PRL messenger RNA and increased expression of the insulin-like growth factor-binding protein-1 messenger RNA. The St-2 cell PRL transcript was larger than the pituitary PRL transcript, suggesting its initiation from the distal, nonpituitary, PRL promoter. This was confirmed by reverse transcription-PCR analysis of PRL transcripts using primers specific for the additional sequences present only in the 5'-untranslated region of RNA initiated from the distal promoter. Transient transfection of a reporter construct containing 3000 bp of DNA 5' to the decidual-specific promoter of the human PRL gene demonstrated that cAMP was capable of activating this distal promoter in St-2 cells. In conclusion, this novel cell line provides an interesting new model in which to pursue aspects of biochemical differentiation of human endometrium in vitro.

Chlamydia trachomatis infection of human endometrial stromal cells induces defective decidualisation and chemokine release

“…To assess the effects of chlamydial infection on the decidualisation of primary endometrial stromal cells, an in-vitro decidualisation method previously described was used 32 . Cells were trypsinised, spun for 3.5 minutes at 800 rpm and resuspended phenol red-free RPMI 1640 with 10% CSFCS.…”

Gene induction and categorical reprogramming during in vitro human endometrial fibroblast decidualization

“…The treated endometrial cells acquire the characteristic morphologic phenotype of decidual cells and express decidual cell-specific markers such as prolactin and insulin-like growth factor binding protein-1 (IGFBP-1). Decidualization can also be induced in primary cultures of human decidual fibroblasts (36) and in a permanent cell line of human endometrial fibroblasts (St-2 cells) (8). Treatment of primary cultures of decidual fibroblasts and St-2 cells with cAMP induces prolactin gene expression and decidualization, but treatment with progesterone, alone or in combination with estradiol, is without effect.…”

N5 endometrial stromal cell line: A model system to study decidual prolactin gene expression

“…Previous studies of prolactin gene expression in endometrial cells have been performed in primary cultures of human endometrial LTo whom correspondence should be addressed at Division of Endocrinology, Children's Hospital Medical Center, 3333 Burner Avenue, Cincinnati, Ohio 45229-3039. stromal cells and in the St-2 endometrial cell line described by White and coworkers (6). Studies in both the primary endometrial stromal cell cultures and the St-2 cell line have shown that the prolactin gene is not expressed until the cells have been exposed to hormones or other factors that induce the cells to decidualize.…”

“…The differences in regulation are due in large part to the fact that decidual prolactin is regulated by an alternative promoter located 5' of an additional noncoding exon (exon la) located approximately 6 kilobases (kb) upstream of the pituitary prolactin transcription initiation site (9,16). The decidual type prolactin promoter is also used by other extrapituitary cells that express protactin, such as lymphocytes (2,6,11), myometrial ceils (3) and cyclic AMP-treated decidual fibroblasts (19) and dermal fibroblasts (20).…”