Human endometrial epithelial cells grown on collagen in serum-free medium. Estrogen responsiveness and morphology

Casslén, Bertil; Harper, M. J. K.

doi:10.1530/acta.0.1250101

Cited by 11 publications

(9 citation statements)

References 15 publications

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“…Recently and in earlier studies human endometrial epithelial cells have been reported to be cultured on rattail collagen type I matrix (Casslen and Harper 1991;Ghosh and Sengupta 1995). It is reported that cells plated on top of collagen I gels deposit their own basement membrane (Emerman and Pitelka 1977;Streuli and Bissell 1990).…”

Section: Influence Of the Extracellular Matrixmentioning

confidence: 98%

Establishment of a human endometrial cell culture system and characterization of its polarized hormone responsive epithelial cells

Claßen-Linke

Kusche

Knauthe³

et al. 1996

Cell and Tissue Research

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Uterine epithelial cells from normal human endometrium were cultured as a primary cell culture in a dual-chambered system. The epithelial cells were isolated from endometrial tissue of the proliferative phase obtained by hysterectomy. The epithelial cells were seeded on Millicell CM filters coated with the extracellular matrix Matrigel. Depending on the culture conditions, the epithelial cells formed a polarized cell monolayer on Matrigel or gland-like structures in Matrigel. The epithelial cell polarity was maintained during culture, which could be proved by electron microscopy. The progesterone and estrogen receptors as typical marker molecules for physiologically intact endometrial epithelial cells could be detected immunohistochemically as well as by RT-PCR in vitro and were down-regulated by medroxyprogesterone acetate (MPA) used as progesterone analogue. As this cell culture system exhibits morphological and immunohistochemical characteristics, typical for the in vivo situation, and since it can be modulated by hormone treatment under the in vitro conditions described, it represents a valuable tool for investigating processes that are essential for endometrial differentiation and reproductive functions.

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Section: Influence Of the Extracellular Matrixmentioning

confidence: 98%

Establishment of a human endometrial cell culture system and characterization of its polarized hormone responsive epithelial cells

Claßen-Linke

Kusche

Knauthe³

et al. 1996

Cell and Tissue Research

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show abstract

“…The methods established in the present study will help to obtain highly purified epithelial cells for subsequent culture. Casslen and Harper [14] were able to culture human endometrial epithelial cells in serum-free medium using collagen coated tissue culture plates. We also used collagen coated culture plates in the present study.…”

Section: Fig 2 Diagram Of Density Gradient Centrifugation Withmentioning

confidence: 99%

“…Several studies have reported the serum-free culture of human [14], murine [15], guinea pig [16] and porcine [12] epithelial cells. However, there have been no studies on serum-free culture of bovine endometrial epithelial cells.…”

mentioning

confidence: 99%

Isolation and Culture of Bovine Endometrial Epithelial Cells in a Serum-Free Culture System.

Takahashi

Iga

Sato

et al. 2001

Journal of Reproduction and Development

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Abstract. The objective of this study was to establish a serum-free culture system for bovine endometrial epithelial cells, and to identify the function of these cells. Epithelial cells were separated from uterine endometrium at Days 5-10 of the estrous cycle by 0.76% EDTA-PBS and collagenase treatment. After histological observation, the epithelium was completely separated from the basement membrane by EDTA treatment. The separated epithelial cells were then dispersed by collagenase and cultured in Dulbecco's Modified Eagle's Medium/Ham's F-12 1:1 supplemented with insulin, transferrin, sodium-selenite, hydrocortisone, retinol, ascorbic acid and antibiotics on collagencoated culture dishes. The cells attached to the substrate and reached confluency after 5-7 days of culture. An immunocytochemical study revealed that the cultured epithelial cells were positive to cytokeratin, but not vimentin. The production of prostaglandin (PG) F2α was significantly higher (p<0.001) in the presence of 100 nM oxytocin (OT; 2.81 fold greater than the concentration in the control) than in the absence of OT. These results indicate that it is possible to culture bovine endometrial epithelial cells in the serum-free culture system, and that cultured cells have some typical characteristics of endometrial epithelial cells.

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“…As uterine receptivity is a polarity-dependent specialized property of endometrial epithelial cells (see above) and is essential for their differentiation, a polarized monolayer of UE cells can be considered as providing conditions equivalent to those in situ (Casslen and Harper 1991). The UE cell monolayer developed on an ECMcoated surface displays distinct polarization with the basal region lying adjacent to the ECM and the apical region being similar to the surface of the cells facing the uterine lumen.…”

Section: Introductionmentioning

confidence: 99%

Demonstration of uterine receptivity in vitro by co-culture of rat epithelial cells and blastocyst

et al. 2006

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Uterine receptivity is prerequisite for the attachment of the embryo to the uterine epithelium and involves a specialized polarity-dependent property of uterine epithelial (UE) cells. These UE cells, when polarized in culture, behave like cells in utero by exhibiting apico-basal polarity. In order to develop an implantation model in vitro, UE cells were polarized on extracellular matrix (ECM), and polarity was validated by response to estradiol-17beta administered exogenously. UE cells of pregnant rats at day-3 and day-4 post-coitum (p.c.) and of non-pregnant rats were cultured on bare and extracellular-matrix-coated petri dishes until confluency. Hatched blastocysts were transferred to the cultures, and adhesion was monitored every 24 h. Although blastocysts attached to UE cells that were taken from non-pregnant rats and from rats of day-3 p.c. and cultured on bare plastic, they failed to attach to these cells polarized on ECM. However, blastocysts attached firmly to UE cells that had been taken from rats of day-4 p.c. and polarized on ECM. Receptivity of UE cells taken from non-pregnant and pregnant (day-4 p.c.) rats was quantitated by flow cytometric estimation of cellular levels of beta3 integrin. The expression of beta3 integrin in UE cells from rats of day-4 p.c. was highly significant (P<0.01) when compared with its expression in UE cells from non-pregnant rats. The expression of beta3 integrin in UE cells of day-4 p.c. confirmed the receptivity of these cells to blastocyst implantation. Uterine receptivity was also validated in vivo by inducing the decidual cell reaction in rats ovariectomized on day-3 and day-4 p.c. Whereas remarkable deciduoma was noticed in the animals of day-4 p.c., it was absent in the animals of day-3 p.c., thereby indicating that the uterus was receptive on day-4 p.c. only. Thus, blastocysts do not attach to polarized UE cells that have been obtained from a non-receptive uterus. Attachment will occur only if the cells are obtained from a receptive uterus. UE cell receptivity is therefore essential for mimicking the process of implantation in vitro.

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Human endometrial epithelial cells grown on collagen in serum-free medium. Estrogen responsiveness and morphology

Abstract: The aim of the study was to explore the possibility of using human endometrial epithelial cells in serum-free culture as a sensitive assay for hormonal effects on the human endometrium. Glands were isolated fol-

Cited by 11 publications

References 15 publications

Establishment of a human endometrial cell culture system and characterization of its polarized hormone responsive epithelial cells

Establishment of a human endometrial cell culture system and characterization of its polarized hormone responsive epithelial cells

Isolation and Culture of Bovine Endometrial Epithelial Cells in a Serum-Free Culture System.

Demonstration of uterine receptivity in vitro by co-culture of rat epithelial cells and blastocyst

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