Isolation and Culture of Human Endometrial Glands*

Satyaswaroop, P.G.; Bressler, Robert S.; Peña, Maria; Gurpide, Erlio

doi:10.1210/jcem-48-4-639

Cited by 209 publications

(53 citation statements)

References 5 publications

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“…The nulliparous patient, aged 37, had undergone surgery because of uterine myomatosis. A modification of the isolation protocol by Satyaswaroop et al (1979) was used. Briefly, several endometrial tissue specimens from the region of the uterine corpus were cut into 1-3 mm 3 pieces, washed in PBS, digested for 45 min at 37 C in PBS with 4 mg/ml BSA (Sigma) containing 2·5 mg/ml collagenase (CLSII, 'Worthington type'; Biochrom, Berlin, Germany) and 25 µg/ml DNAse (Sigma) and passed through a 250 µm sieve to remove mucous material and undigested tissue.…”

Section: Isolation and Immortalization Of Human Eecsmentioning

confidence: 99%

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Hombach‐Klonisch¹,

Kehlen²,

Fowler³

et al. 2005

Journal of Molecular Endocrinology

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Information on the regulation of steroid hormone receptors and their distinct functions within the human endometrial epithelium is largely unavailable. We have immortalized human primary endometrial epithelial cells (EECs) isolated from a normal proliferative phase endometrium by stably transfecting the catalytic subunit (hTERT) of the human telomerase complex and cultured these hTERT-EECs now for over 350 population doublings. Active hTERT was detected in hTERT-EECs employing the telomerase repeat amplification assay protocol. hTERT-EECs revealed a polarized, non-invasive epithelial phenotype with apical microvilli and production of a basal lamina when grown on a three-dimensional collagen-fibroblast lattice. Employing atomic force microscopy, living hTERT-EECs were shown to produce extracellular matrix (ECM) components and ECM secretion was modified by estrogen and progesterone (P4). hTERT-EECs expressed inducible and functional endogenous estrogen receptor-alpha (ER-alpha) as demonstrated by estrogen response element reporter assays and induction of P4 receptor (PR). P4 treatment down-regulated PR expression, induced MUC-1 gene activity and resulted in increased ER-beta transcriptional activity. Gene activities of cytokines and their receptors interleukin (IL)-6, leukemia inhibitory factor (LIF), IL-11 and IL-6 receptor (IL6-R), LIF receptor and gp130 relevant to implantation revealed a 17 beta-estradiol (E2)-mediated up-regulation of IL-6 and an E2-and P4-mediated up-regulation of IL6-R in hTERT-EECs. Thus, hTERT-EECs may be regarded as a novel in vitro model to investigate the role of human EECs in steroid hormone-dependent normal physiology and pathologies, including implantation failure, endometriosis and endometrial cancer.

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Section: Isolation and Immortalization Of Human Eecsmentioning

confidence: 99%

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Hombach‐Klonisch¹,

Kehlen²,

Fowler³

et al. 2005

Journal of Molecular Endocrinology

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“…Endometrial tissue was washed thoroughly with physiological saline, minced with scissors and enzymatically dispersed with collagenase (2 g/l) and deoxyribonuclease (DNase, 50 mg/l). Endometrial stromal cells were separated from epithelial cells by filtration through 38 mm steel mesh, as described previously (15). The cells were suspended in R.P.M.I-1640 containing 10% FCS in plastic T flasks (75 cm 2 , Falcon, NJ, USA) in a humidified 37 ЊC atmosphere of 5% CO 2 -95% air.…”

Section: Cell Culture Of Human Endometrial Stromal Cellsmentioning

confidence: 99%

Relationship between the release of prolactin and endothelin-1 in human decidualized endometrial cells

Kubota

Taguchi²,

Kobayashi³

et al. 1997

European Journal of Endocrinology

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The study was undertaken to investigate the interaction between the release of endothelin-1 (ET-1) and prolactin (PRL) from human decidualized endometrial cells, and the effects of transforming growth factor (TGF)-b1 on the release of ET-1 and PRL from human decidual cells in the early stage of pregnancy. Stromal cells derived from human endometrial tissues were cultured on a type-1 collagen membrane in serum-free medium with or without 100 nmol/l oestradiol and 1 mmol/l medroxyprogesterone acetate (MPA) for 12 days. In addition, decidual cells of early pregnancy were cultured with or without TGF-b1 for 48 h. PRL and ET-1 levels in the media were measured by a specific enzyme immunoassay and RIA respectively.In decidualized endometrial cells induced by ovarian steroid hormones in vitro, PRL release increased and ET-1 release decreased in a time-dependent manner. Ovarian steroid hormones significantly attenuated ET-1 release. In the decidual cells of early pregnancy, TGF-b1 significantly attenuated the PRL release, whereas this growth factor dose-dependently increased ET-1 release. There was a significant negative correlation between the release of PRL and that of ET-1.These results demonstrate that the regulation of PRL release is quite different from that of ET-1 release in the human decidualized endometrial cells, and suggest that TGF-b1 has significant effects on PRL and ET-1 release in the human decidua of early pregnancy.

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“…Normal tissue samples at the proliferative stage were minced finely with scissors and washed in phosphate buffered saline. The endometrial stromal cells were then isolated as described previously [17]. Briefly, the minced tissue samples were digested in calcium-and magnesium-free Hank's solution (CMF-Hanks) containing 0.25% collagenase (Type IA; Sigma-Aldrich, St. Louis, MO, USA) and then strained through a 250-µm stainless steel sieve (Sanpo, Tokyo, Japan) to remove undigested tissue and mucous material.…”

Section: Tissue Collection and Immortalization Of Endometrial Stromalmentioning

confidence: 99%

Involvement of Stathmin in Proliferation and Differentiation of Immortalized Human Endometrial Stromal Cells

Tamura

Yoshie

Hara

et al. 2007

Journal of Reproduction and Development

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Abstract. Uterine endometrial stromal cells differentiate into decidual cells during the late secretory phase of the menstrual cycle and pregnancy. However, the biochemical mechanisms of decidualization have yet to be definitively elucidated. In the present study, we transfected primary human endometrial stromal cell with a temperature-sensitive mutant of simian virus 40 large T antigen and thereby established an immortalized stromal cell line (EtsT) in order to examine the role of stathmin, a cytosolic phosphoprotein that regulates microtubule dynamics, in stromal cell differentiation. When treated with the decidual stimulus dibutyryl-cAMP (db-cAMP) or forskolin, the fibroblastic cell-shaped EtsT cells transformed into large-and round-shaped cells and secreted large amounts of the decidual markers prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP-1). Analysis of the stathmin protein levels in the db-cAMP-and forskolin-treated EtsT cells revealed that the total and phosphorylated protein levels dropped as decidualization progressed. Suppression of stathmin expression by transfection with small interfering RNA (siRNA) suppressed EtsT cell proliferation. It also abolished db-cAMP-induced PRL and IGFBP-1 mRNA expression and protein secretion. Thus, stathmin expression can be considered an integral factor regulating the initial stage of the process of human endometrial stromal cell differentiation.

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Isolation and Culture of Human Endometrial Glands*

Cited by 209 publications

References 5 publications

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Relationship between the release of prolactin and endothelin-1 in human decidualized endometrial cells

Involvement of Stathmin in Proliferation and Differentiation of Immortalized Human Endometrial Stromal Cells

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