scholarly journals Reporters to mark and eliminate basal or luminal epithelial cells in culture and in vivo

PLoS Biology ◽  
2018 ◽  
Vol 16 (6) ◽  
pp. e2004049 ◽  
Author(s):  
Olmo Sonzogni ◽  
Jennifer Haynes ◽  
Laurie A. Seifried ◽  
Yahia M. Kamel ◽  
Kai Huang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cells In Culture ◽  
Luminal Epithelial Cells

scholarly journals DAAM mediates the assembly of long-lived, treadmilling stress fibers in collectively migrating epithelial cells in Drosophila

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Kristin M Sherrard ◽  
Maureen Cetera ◽  
Sally Horne-Badovinac
Keyword(s):  
Epithelial Cells ◽  
Cell Movement ◽  
Stress Fibers ◽  
Linear Trajectory ◽  
Epithelial Migration ◽  
Cells In Culture ◽  
Multiple Cell ◽  
Developmental Switch ◽  
Over Time

Stress fibers (SFs) are actomyosin bundles commonly found in individually migrating cells in culture. However, whether and how cells use SFs to migrate in vivo or collectively is largely unknown. Studying the collective migration of the follicular epithelial cells in Drosophila, we found that the SFs in these cells show a novel treadmilling behavior that allows them to persist as the cells migrate over multiple cell lengths. Treadmilling SFs grow at their fronts by adding new integrin-based adhesions and actomyosin segments over time. This causes the SFs to have many internal adhesions along their lengths, instead of adhesions only at the ends. The front-forming adhesions remain stationary relative to the substrate and typically disassemble as the cell rear approaches. By contrast, a different type of adhesion forms at the SF’s terminus that slides with the cell’s trailing edge as the actomyosin ahead of it shortens. We further show that SF treadmilling depends on cell movement and identify a developmental switch in the formins that mediate SF assembly, with Dishevelled-associated activator of morphogenesis acting during migratory stages and Diaphanous acting during postmigratory stages. We propose that treadmilling SFs keep each cell on a linear trajectory, thereby promoting the collective motility required for epithelial migration.

scholarly journals Differential Effects of Gamma Interferon on Chlamydia trachomatis Growth in Polarized and Nonpolarized Human Epithelial Cells in Culture

1998 ◽  
Vol 66 (5) ◽  
pp. 2349-2351 ◽  
Author(s):  
Colleen D. Kane ◽  
Gerald I. Byrne
Keyword(s):  
Chlamydia Trachomatis ◽  
Epithelial Cells ◽  
Gamma Interferon ◽  
Growth Modulation ◽  
Differential Effects ◽  
Cells In Culture ◽  
Human Epithelial Cells ◽  
Polarized Cells ◽  
Ifn Γ

ABSTRACT The effects of gamma interferon (IFN-γ) on Chlamydia trachomatis growth in polarized epithelial cells were examined. The range of IFN-γ concentrations causing aberrant chlamydial growth was wider in polarized than in nonpolarized cultures. Results indicate that chlamydial growth modulation in polarized cells readily leads to persistence and better reflects in vivo conditions.

scholarly journals Normal and tumor-derived myoepithelial cells differ in their ability to interact with luminal breast epithelial cells for polarity and basement membrane deposition

2002 ◽  
Vol 115 (1) ◽  
pp. 39-50 ◽  
Author(s):  
Thorarinn Gudjonsson ◽  
Lone Rønnov-Jessen ◽  
René Villadsen ◽  
Fritz Rank ◽  
Mina J. Bissell ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Specific Antigen ◽  
Myoepithelial Cells ◽  
Normal Breast ◽  
Breast Epithelial ◽  
Luminal Epithelial Cells ◽  
Laminin 1

The signals that determine the correct polarity of breast epithelial structures in vivo are not understood. We have shown previously that luminal epithelial cells can be polarized when cultured within a reconstituted basement membrane gel. We reasoned that such cues in vivo may be given by myoepithelial cells. Accordingly, we used an assay where luminal epithelial cells are incorrectly polarized to test this hypothesis. We show that culturing human primary luminal epithelial cells within collagen-I gels leads to formation of structures with no lumina and with reverse polarity as judged by dual stainings for sialomucin, epithelial specific antigen or occludin. No basement membrane is deposited, and β4-integrin staining is negative. Addition of purified human myoepithelial cells isolated from normal glands corrects the inverse polarity, and leads to formation of double-layered acini with central lumina. Among the laminins present in the human breast basement membrane (laminin-1, -5 and -10/11), laminin-1 was unique in its ability to substitute for myoepithelial cells in polarity reversal.Myoepithelial cells were purified also from four different breast cancer sources including a biphasic cell line. Three out of four samples either totally lacked the ability to interact with luminal epithelial cells, or conveyed only correction of polarity in a fraction of acini. This behavior was directly related to the ability of the tumor myoepithelial cells to produce α-1 chain of laminin. In vivo, breast carcinomas were either negative for laminin-1 (7/12 biopsies) or showed a focal, fragmented deposition of a less intensely stained basement membrane (5/12 biopsies). Dual staining with myoepithelial markers revealed that tumor-associated myoepithelial cells were either negative or weakly positive for expression of laminin-1, establishing a strong correlation between loss of laminin-1 and breast cancer. We conclude that the double-layered breast acinus may be recapitulated in culture and that one reason for the ability of myoepithelial cells to induce polarity is because they are the only source of laminin-1 in the breast in vivo. A further conclusion is that a majority of tumor-derived/-associated myoepithelial cells are deficient in their ability to impart polarity because they have lost their ability to synthesize sufficient or functional laminin-1. These results have important implications for the role of myoepithelial cells in maintenance of polarity in normal breast and how they may function as structural tumor suppressors.

Progesterone-dependent and -independent modulation of luminal epithelial transcription to support pregnancy in cattle

2021 ◽  
Author(s):  
Thiago Martins ◽  
Mariana Sponchiado ◽  
Felipe Alves Correa Carvalho Silva ◽  
Eliab Estrada-Cortés ◽  
Peter J. Hansen ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Embryo Transfer ◽  
Pregnancy Outcomes ◽  
Biosynthetic Activity ◽  
Phosphoinositide Signaling ◽  
Uterine Environment ◽  
Luminal Cells ◽  
Luminal Epithelial Cells ◽  
Layer Control

In cattle, starting 4-5 days after estrus, pre-implantation embryonic development occurs in the confinement of the uterine lumen. Cells in the endometrial epithelial layer control the molecular traffic to and from the lumen and, thereby determine luminal composition. Starting early post-estrus, endometrial function is regulated by sex-steroids, but the effects of progesterone on luminal cells transcription have not been measured in vivo. First objective was to determine the extent to which progesterone controls transcription in luminal epithelial cells 4 d (D4) after estrus. Second objective was to discover luminal transcripts that predict pregnancy outcomes, when the effect of progesterone is controlled. Endometrial luminal epithelial cells were collected from embryo transfer recipients on D4 using a cytological brush and their transcriptome determined by RNASeq. Pregnancy by embryo transfer was measured on D30 (25 pregnant and 18 non-pregnant). Progesterone concentration on D4 was associated positively (n= 182) and negatively (n= 58) with gene expression. Progesterone-modulated transcription indicated an increase in oxidative phosphorylation, biosynthetic activity and proliferation of epithelial cells. When these effects of progesterone were controlled, different genes affected positively (n= 22) and negatively (n= 292) odds of pregnancy. These set of genes indicated that a receptive uterine environment was characterized by the inhibition of phosphoinositide signaling and innate immune system responses. A panel of 25 genes predicted the pregnancy outcome with sensitivity and specificity ranging from 64-96% and 44-83%, respectively. In conclusion, in the early diestrus, both progesterone-dependent and -independent mechanisms regulate luminal epithelial transcription associated with pregnancy outcomes in cattle.

scholarly journals Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

2020 ◽  
Vol 2020 ◽  
pp. 1-15 ◽  
Author(s):  
Katarzyna Wojtanowicz-Markiewicz ◽  
Magdalena Kulus ◽  
Sandra Knap ◽  
Ievgenia Kocherova ◽  
Maurycy Jankowski ◽  
...  
Keyword(s):  
Cell Culture ◽  
Epithelial Cells ◽  
Primary Cell Culture ◽  
Water Movement ◽  
Inorganic Ions ◽  
Primary Cell ◽  
Lag Phase ◽  
Luminal Epithelial Cells

Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx’s behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.

Prokineticin 1–prokineticin receptor 1 signaling promotes angiogenesis in the porcine endometrium during pregnancy†

2020 ◽  
Vol 103 (3) ◽  
pp. 654-668 ◽  
Author(s):  
Ewelina Goryszewska ◽  
Piotr Kaczynski ◽  
Gianfranco Balboni ◽  
Agnieszka Waclawik
Keyword(s):  
Epithelial Cells ◽  
Reproductive Tract ◽  
Secretory Protein ◽  
Angiogenic Factor ◽  
In Vivo Model ◽  
Secretory Function ◽  
Pleiotropic Functions ◽  
Estradiol 17Β ◽  
Luminal Epithelial Cells

Abstract Pregnancy establishment in mammals, including pigs, requires proper communication between embryos and the maternal reproductive tract. Prokineticin 1 (PROK1) has been described as a secretory protein with pleiotropic functions and as a novel tissue-specific angiogenic factor. However, despite the studies performed mainly on human cell lines and in mice, the function of PROK1 in the endometrium during early pregnancy is still not fully elucidated. We hypothesized that PROK1 contributes to pregnancy establishment in pigs. The present study is the first to report that the expression of PROK1 and its receptor (PROKR1) is elevated in the porcine endometrium during the implantation and early placentation period. PROK1 protein was detected mainly in luminal epithelial cells, glandular epithelial cells, and blood vessels in the endometrium. Using the porcine in vivo model of unilateral pregnancy, we revealed that conceptuses induced the endometrial expression of PROK1 and PROKR1. Moreover, the embryonic signal, estradiol-17β, as well as progesterone, stimulated the endometrial expression of PROK1 and PROKR1. We also evidenced that PROK1–PROKR1 signaling supports endometrial angiogenesis in pigs. The PROK1-stimulated proliferation of primary porcine endometrial endothelial (PEE) cells involved PI3K/AKT/mTOR, MAPK, cAMP, and NFKB signaling pathways. Furthermore, PROK1 via PROKR1 promoted the formation of capillary-like structures by PEE cells. PROK1 also stimulated VEGFA and PGF2α secretion, which in turn may indirectly support angiogenic changes within endometrial tissue. In summary, our study suggests that PROK1 acts as an embryonic signal mediator that regulates endometrial angiogenesis and secretory function during the implantation and early placentation period in pigs.

Anti-estrogenic effect of TAS-108 in breast cancer cell lines.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. e11029-e11029
Author(s):  
Hadong Kim ◽  
Young Choi ◽  
Tae-Hoon Kim
Keyword(s):  
Breast Cancer ◽  
Epithelial Cells ◽  
Growth Inhibition ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cell Lines ◽  
Luminal Epithelial Cells ◽  
Mcf 7

e11029 Background: TAS-108 (TAS) is an antiestrogen that binds to both ERα and ERβ and inhibits both estrogen-dependent and independent tumors and TAM resistant tumors. Breast cancer cell lines (BCC) with different pheno- and genotypes are reasonably acceptable models of in vivo breast cancer cell behavior. Thus, we have investigated anti-estrogenic activity of TAS on BCC in the presence of diarylpropionitrile (DPN), a pure ERβ agonist to find ERβ role in TAS activity. Methods: Cultured cells of (1) MCF-7 and ZR-75 (luminal epithelial cells), (2) BT-474 and SK-BR-3 (weakly luminal epithelial cells), and (3) MDA-MB-231 and Hs578T (invasive, mesenchymal-like cells) were treated with: (1) increasing concentrations of DPN or E2, (2) 100nM of 4-hydroxytamoxifen (4-HT) or TAS, and (3) 100nM of DPN or E2 with increasing concentrations of either 4-HT or TAS. After 6 days, cell proliferation was analyzed. The mRNA levels of ERβ and ERα isoform were determined by RT-qPCR. ERα, PR, and Her-2 status was determined by IHC. Percent proliferation compared to the control was calculated and P <0.05 based on T-test was considered significant. Results: The most significant proliferation and growth inhibition was seen in MCF-7 BCC. In the presence of DPN, IC50 (50% inhibitory concentration) was 12.5nmol of TAS vs 33.1nmol of 4-HT. In the presence of E2, IC50 was 2.15 nmol of TAS vs 49.7nmol of 4-HT. Other BCC showed insignificant or variable proliferative and inhibitory response. Overall, DPN displayed high proliferation in most BCC. TAS exhibited higher growth inhibition than 4-HT. Conclusions: Variable responses of different BCC lines to ERα and ERβ agonists and antagonists may be instructive in modeling in vivo breast cancer response to these compounds. Overall, exposure to ERβ agonists showed a significant proliferation in ERα+, ERα- and ERβ+ cells. TAS activity was more pronounced and its anti-estrogen activity may involve ERβ in addition to ERα. [Table: see text]

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