Characterization of a Polarized Porcine Uterine Epithelial Model System1

Bowen, Jeffery A.; Newton, G. R.; Weise, D W; Bazer, Fuller W.; Burghardt, Robert C.

doi:10.1095/biolreprod55.3.613

Cited by 31 publications

(14 citation statements)

References 24 publications

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“…This is in contrast to a previous report, where polarised bovine epithelial cells treated with OT had predominantly basolateral accumulation of PGF, but not PGE (Asselin et al 1996). However, porcine endometrial epithelial cells and the bovine endometrial epithelial cell line, bEEL, reportedly have predominantly basolateral release of both PGE and PGF (Bowen et al 1996, Lacroix-Pepin et al 2011. Prostaglandins are charged anions that diffuse poorly across cell membranes, thus require facilitated transport by the prostaglandin transporter (PGT) and multidrug resistance-associated protein 4 (MRP4; Schuster 1998, Banu et al 2003, Tithof et al 2007, Lacroix-Pepin et al 2011.…”

Section: Discussioncontrasting

confidence: 99%

Polarised bovine endometrial epithelial cells vectorially secrete prostaglandins and chemotactic factors under physiological and pathological conditions

MacKintosh¹,

Schuberth²,

Healy³

et al. 2013

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Epithelial cells of the endometrium secrete prostaglandins to regulate the bovine oestrous cycle and form a functional barrier to microbes. However, bacterial infection of the endometrium commonly causes infertility in dairy cattle by disrupting endometrial physiology. Epithelial cell cultures are used to study the mechanisms of physiology and pathology, but 2D cultures may not reflect the 3D complexity of the epithelium. In this study, a polarised epithelial cell transwell culture was developed, using transepithelial resistance (TER), to monitor epithelial integrity. Polarised epithelial cells were treated with oxytocin and arachidonic acid to test physiological function and with lipopolysaccharide (LPS) to mimic bacterial infection. Supernatants were analysed for prostaglandin E 2 (PGE), prostaglandin F 2a , the chemokine interleukin-8 (IL8) and the ability of supernatants to induce neutrophil migration. Confluent epithelial cells established polarity when TER was O1800 Ucm 2 and predominantly released prostaglandins basolaterally. In contrast, IL8 from epithelial cells accumulated apically and the supernatants were highly chemotactic for neutrophils. The striking exception was when the epithelial cells were treated with LPS in the apical or basolateral compartment independently, which led to the release of IL8 towards the treated compartment. Although stromal cells also accumulated PGE and IL8 in response to treatment, co-culture of stromal cells in the well below polarised epithelial cells did not influence cellular responses. In conclusion, polarised endometrial epithelial cells vectorially released prostaglandins and chemokines to reflect their respective mechanistic roles in physiology and pathology.

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Section: Discussioncontrasting

confidence: 99%

Polarised bovine endometrial epithelial cells vectorially secrete prostaglandins and chemotactic factors under physiological and pathological conditions

MacKintosh¹,

Schuberth²,

Healy³

et al. 2013

Reproduction

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“…Luminal epithelial cells, glandular epithelial cells, and stromal cells were isolated aseptically and separated from the three different sites of the genital tract using a combination and some modifications of procedures previously described (5,6,28,31,32).…”

Section: Methodsmentioning

confidence: 99%

“…In addition, polarized porcine epithelial cell model systems have been developed for in vitro analysis of pregnancy paradigms, actions of hormones, prostaglandin secretion, and epithelial cell-stromal cell interactions (5,6,24,32).…”

mentioning

confidence: 99%

Primary Cultures of Female Swine Genital Epithelial Cells In Vitro: a New Approach for the Study of Hormonal Modulation of Chlamydia Infection

et al. 2003

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Previous studies have demonstrated that female reproductive hormones influence chlamydial infection both in vivo and in vitro. Due to the reduced availability of human genital tissues for research purposes, an alternative hormone-responsive model system was sought to study chlamydial pathogenesis. Mature female swine eliminated from breeding programs were selected as the animals of choice because of the similarity of a sexually transmitted disease syndrome and sequelae in swine to a disease syndrome and sequelae found in humans, because of the near identity of a natural infectious chlamydial isolate from swine to Chlamydia trachomatis serovar D from humans, and because a pig's epithelial cell physiology and the mean length of its estrous cycle are similar to those in humans. Epithelial cells from the cervix, uterus, and horns of the uterus were isolated, cultivated in vitro in Dulbecco's minimum essential medium-Hanks' F-12 (DMEM-F-12) medium with and without exogenous hormone supplementation, and analyzed for Chlamydia suis S-45 infectivity. The distribution of chlamydial inclusions in swine epithelial cells was uneven and was influenced by the genital tract site and hormone status. This study confirmed that, like primary human endometrial epithelial cells, estrogen-dominant swine epithelial cells are more susceptible to chlamydial infection than are progesterone-dominant cells. Further, the more differentiated luminal epithelial cells were more susceptible to infection than were glandular epithelial cells. Interestingly, chlamydial growth in mature luminal epithelia was morphologically more active than in glandular epithelia, where persistent chlamydial forms predominated. Attempts to reprogram epithelial cell physiology and thereby susceptibility to chlamydial infection by reversestage, exogenous hormonal supplementation were unsuccessful. Freshly isolated primary pig epithelial cells frozen at ؊80°C in DMEM-F-12 medium with 10% dimethyl sulfoxide for several weeks can, after thawing, reform characteristic polarized monolayers in 3 to 5 days. Thus, primary swine genital epithelia cultured ex vivo appear to be an excellent cell model for dissecting the hormonal modulation of several aspects of chlamydial pathogenesis and infection.

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“…Isolation and characterization of porcine uterine luminal epithelial cells were performed following pancreatindispase enzymatic release of epithelium from hysterectomized gilts as described by Bowen et al (1996). Isolation of uterine epithelium from equine endometrial biopsies was performed as described by Brady et al (1993).…”

Section: Isolation Of Uterine Endometrial Epithelial Cellsmentioning

confidence: 99%

Endometrial connexin expression in the mare and pig: Evidence for the suppression of cell-cell communication in uterine luminal epithelium

et al. 1998

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This investigation examines the relationship between implantation strategy and gap junction protein expression in uterine endometrium. The pattern of gap junction and connexin protein expression was analyzed in porcine and equine endometrium from cycling and pregnant animals using electron microscopy and immunocytochemistry. Functional analysis of cellcell communication was also monitored by laser cytometry in primary cultures of endometrial epithelial cells. Gap junctions were detected in endometrial stroma of cycling and pregnant animals, which was correlated with immunoreactive Cx43 within stromal fibroblasts and vascular elements. No Cx26, Cx32, or Cx43 immunostaining was detected in luminal endometrial epithelium in either the mare or the pig at any stage of the estrous cycle or pregnancy. In contrast, endometrial glands of the mare exhibited a spatiotemporal pattern of Cx43 expression in the apicolateral plasma membrane which, when present, colocalized with the tight junction-associated protein, ZO-1. Uterine glandular Cx43 expression in mares was present from day 3 postovulation through day 14 of diestrus and until day 23 of pregnancy, whereas Cx43 was absent within uterine glands during seasonal anestrus, estrus, and after day 30 of pregnancy. Primary cultures of equine endometrial epithelial cells expressed both immunoreactive Cx43 and significant gap junction-mediated intercellular communication (GJIC) which was rapidly upregulated by 1.0 mM 8-bromo-cAMP or blocked with 1.0 mM octanol. No GJIC or connexin protein was detected in cultured porcine epithelial cells despite incubation with a variety of agents, including 8-bromo-cAMP, steroid hormones, retinoic acid, and/or prolactin.

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Characterization of a Polarized Porcine Uterine Epithelial Model System1

Cited by 31 publications

References 24 publications

Polarised bovine endometrial epithelial cells vectorially secrete prostaglandins and chemotactic factors under physiological and pathological conditions

Polarised bovine endometrial epithelial cells vectorially secrete prostaglandins and chemotactic factors under physiological and pathological conditions

Primary Cultures of Female Swine Genital Epithelial Cells In Vitro: a New Approach for the Study of Hormonal Modulation of Chlamydia Infection

Endometrial connexin expression in the mare and pig: Evidence for the suppression of cell-cell communication in uterine luminal epithelium

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