scholarly journals Evaluating the Glucantime Concentration for the ex vivo Glial Cell Model of Antimony-resistant Leishmania tropica Amastigotes

2021 ◽  
Vol 45 (4) ◽  
pp. 237-240
Author(s):  
Orçun Zorbozan ◽  
Vedat Evren ◽  
Mehmet Harman ◽  
Ahmet Özbilgin ◽  
Özlem Alkan Yılmaz ◽  
...  
Keyword(s):  
Glial Cell ◽  
Cell Model ◽  
Ex Vivo ◽  
Leishmania Tropica

scholarly journals Development of a Novel ex vivo Nasal Epithelial Cell Model Supporting Colonization With Human Nasal Microbiota

2019 ◽  
Vol 9 ◽  
Author(s):  
Derald D. Charles ◽  
James R. Fisher ◽  
Sarah M. Hoskinson ◽  
Audrie A. Medina-Colorado ◽  
Yi C. Shen ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Model ◽  
Ex Vivo ◽  
Nasal Epithelial Cell ◽  
Nasal Microbiota

scholarly journals si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Limin Liu ◽  
Guobin Chen ◽  
Taoliang Chen ◽  
Wenjuan Shi ◽  
Haiyan Hu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Stromal Cells ◽  
Target Genes ◽  
Cell Model ◽  
Ex Vivo ◽  
Signalling Pathway ◽  
Endometrial Stromal Cells ◽  
Downstream Target ◽  
Related Proteins ◽  
Tgf Β1

Abstract Background Intrauterine adhesions (IUAs) are manifestations of endometrial fibrosis characterized by inflammation and fibrinogen aggregation in the extracellular matrix (ECM). The available therapeutic interventions for IUA are insufficiently effective in the clinical setting for postoperative adhesion recurrence and infertility problems. In this study, we investigated whether si-SNHG5-FOXF2 can serve as a molecular mechanism for the inhibition of IUA fibrosis ex vivo. Methods FOXF2, TGF-β1 and collagen expression levels were measured by microarray sequencing analysis in three normal endometrium groups and six IUA patients. We induced primary human endometrial stromal cells (HESCs) into myofibroblasts (MFs) to develop an IUA cell model with various concentrations of TGF-β1 at various times. Downstream target genes of FOXF2 were screened by chromatin immunoprecipitation combined with whole-genome high-throughput sequencing (ChIP-seq). We investigated ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related proteins in primary HESCs with FOXF2 downregulation by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting (WB), immunohistochemistry (IHC), flow cytometry, ethylenediurea (EdU) and CCK8 assays. We identified long noncoding RNAs (lncRNA) SNHG5 as the upstream regulatory gene of FOXF2 through RNA immunoprecipitation (RIP), RNA pulldown and fluorescence in situ hybridization (FISH). Finally, we examined FOXF2 expression, ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related proteins in primary HESCs upon FOXF2 downregulation. Results FOXF2 was highly expressed in the endometrium of patients with IUA. Treatment of primary HESCs with 10 ng/ml TGF-β1 for 72 h was found to be most effective for developing an IUA cell model. FOXF2 regulated multiple downstream target genes, including collagen, vimentin (VIM) and cyclin D2/DK4, by ChIP-seq and ChIP-PCR. FOXF2 downregulation inhibited TGF-β1-mediated primary HESC fibrosis, including ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related protein expression. We identified lncRNA SNHG5 as an upstream gene that directly regulates FOXF2 by RIP-seq, qRT-PCR, WB and FISH. SNHG5 downregulation suppressed FOXF2 expression in the IUA cell model, resulting in synergistic repression of the Wnt/β-catenin pathway, thereby altering TGF-β1-mediated ECM aggregation in endometrial stromal cells ex vivo. Conclusions Regulation of the Wnt/β-catenin signalling pathway and ECM formation by si-SNHG5-FOXF2 effectively inhibited the profibrotic effect of TGF-β1 on primary HESCs. This finding can provide a molecular basis for antagonizing TGF-β1-mediated fibrosis in primary HESCs.

scholarly journals Sufu regulation of Hedgehog signaling in P19 cells is required for proper glial cell differentiation

2021 ◽  
Author(s):  
Danielle M. Spice ◽  
Joshua Dierolf ◽  
Gregory M. Kelly
Keyword(s):  
Retinoic Acid ◽  
Cell Differentiation ◽  
Glial Cell ◽  
Neural Differentiation ◽  
Hedgehog Signaling ◽  
Target Genes ◽  
Cell Model ◽  
Ectopic Expression ◽  
Negative Regulator ◽  
Embryonal Carcinoma Cell

AbstractHedgehog signaling is essential for vertebrate development, however, less is known about the negative regulators that influence this pathway during the differentiation of cell fates. Using the mouse P19 embryonal carcinoma cell model, Suppressor of Fused (SUFU), a negative regulator of the Hedgehog pathway, was investigated during retinoic acid-induced neural differentiation. We found Hedgehog signaling was activated in the early phase of neural differentiation and became inactive during terminal differentiation of neurons and astrocytes. SUFU, which regulates signaling at the level of GLI, remained relatively unchanged during the differentiation process, however SUFU loss through CRISPIR-Cas9 gene editing resulted in decreased cell proliferation and ectopic expression of Hedgehog target genes. Interestingly, SUFU-deficient cells were unable to differentiate in the absence of retinoic acid, but when differentiated in its presence they showed delayed and decreased astrocyte differentiation; neuron differentiation did not appear to be affected. Retinoic acid-induced differentiation also caused ectopic activation of Hh target genes in SUFU-deficient cells and while the absence of the GLI3 transcriptional inhibitor suggested the pathway was active, no full-length GLI3 was detected even though the message encoding Gli3 was present. Thus, the study would indicate the proper timing and proportion of glial cell differentiation requires SUFU, and its normal regulation of GLI3 to maintain Hh signaling in an inactive state.

scholarly journals Caveolins in glial cell model systems: from detection to significance

2007 ◽  
Vol 103 (s1) ◽  
pp. 101-112 ◽  
Author(s):  
W. I. Silva ◽  
H. M. Maldonado ◽  
G. Velázquez ◽  
J. O. García ◽  
F. A. González
Keyword(s):  
Glial Cell ◽  
Cell Model ◽  
Model Systems

scholarly journals Functional and anatomical evidence of cerebral tissue hypoxia in young sickle cell anemia mice

2016 ◽  
Vol 37 (3) ◽  
pp. 994-1005 ◽  
Author(s):  
Lindsay S Cahill ◽  
Lisa M Gazdzinski ◽  
Albert KY Tsui ◽  
Yu-Qing Zhou ◽  
Sharon Portnoy ◽  
...  
Keyword(s):  
Blood Flow ◽  
Cerebral Blood Flow ◽  
Brain Tissue ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Cell Model ◽  
Ex Vivo ◽  
Tissue Hypoxia ◽  
Brain Physiology ◽  
The Brain

Cerebral ischemia is a significant source of morbidity in children with sickle cell anemia; however, the mechanism of injury is poorly understood. Increased cerebral blood flow and low hemoglobin levels in children with sickle cell anemia are associated with increased stroke risk, suggesting that anemia-induced tissue hypoxia may be an important factor contributing to subsequent morbidity. To better understand the pathophysiology of brain injury, brain physiology and morphology were characterized in a transgenic mouse model, the Townes sickle cell model. Relative to age-matched controls, sickle cell anemia mice demonstrated: (1) decreased brain tissue pO2 and increased expression of hypoxia signaling protein in the perivascular regions of the cerebral cortex; (2) elevated basal cerebral blood flow , consistent with adaptation to anemia-induced tissue hypoxia; (3) significant reduction in cerebrovascular blood flow reactivity to a hypercapnic challenge; (4) increased diameter of the carotid artery; and (5) significant volume changes in white and gray matter regions in the brain, as assessed by ex vivo magnetic resonance imaging. Collectively, these findings support the hypothesis that brain tissue hypoxia contributes to adaptive physiological and anatomic changes in Townes sickle cell mice. These findings may help define the pathophysiology for stroke in children with sickle cell anemia.

scholarly journals Elevated prolactin redirects secretory vesicle traffic in rabbit lacrimal acinar cells

2007 ◽  
Vol 292 (4) ◽  
pp. E1122-E1134 ◽  
Author(s):  
Yanru Wang ◽  
Christopher T. Chiu ◽  
Tamako Nakamura ◽  
Ameae M. Walker ◽  
Barbara Petridou ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Cell Model ◽  
Ex Vivo ◽  
Ambient Medium ◽  
Adenovirus Vector ◽  
Fluid Secretion ◽  
Secretory Protein ◽  
Apical Plasma Membrane ◽  
Secretory Proteins ◽  
Ductal Epithelial Cell

During pregnancy, lymphocytes infiltrating the rabbit lacrimal gland disperse to the interacinar space from their normal focal concentrations, basal fluid secretion decreases, pilocarpine-induced fluid secretion increases, and stimulated fluid protein concentration decreases. Ductal epithelial cell prolactin (PRL) content increases and redistributes from the apical to the basal-lateral cytoplasm. A replication-incompetent adenovirus vector for rabbit PRL (AdPRL) was used to test the hypothesis that increased intracrine/autocrine PRL signaling alters secretory protein traffic in an ex vivo lacrimal acinar cell model. AdPRL had no discernable influence on microtubules or actin microfilaments or their responses to carbachol (CCh). Endogenous and transduced PRLs exhibited similar, nonpolarized, punctate distributions. Cells secreted PRL consititutively and at increased rates in response to CCh. In contrast, constitutive secretion of β-hexosaminidase was negligible, suggesting that the constitutive pathway for PRL is relatively inaccessible to typical secretory proteins. AdPRL had no significant effect on total secretion of β-hexosaminidase or syncollin-green fluorescent protein (GFP), a chimeric secretory protein construct. However, it reversed the polarized distributions of vesicles containing rab3D and syncollin-GFP. Live-cell imaging indicated that AdPRL redirected CCh-dependent syncollin-GFP exocytosis from the apical plasma membrane to the basal-lateral membrane. Elevated concentrations of exogenous rabbit PRL in the ambient medium elicited similar changes. These observations suggest that elevated PRL, as occurs in the physiological hyperprolactinemia of pregnancy, induces lacrimal epithelial cells to express a mixed exocrine/endocrine phenotype that secretes fluid to the acinus-duct lumen but secretes proteins to the underlying tissue space. This phenotype may contribute to the pregnancy-associated immunoarchitecture.

Altered traffic to the lysosome in an ex vivo lacrimal acinar cell model for chronic muscarinic receptor stimulation

2004 ◽  
Vol 79 (5) ◽  
pp. 665-675 ◽  
Author(s):  
Limin Qian ◽  
Jiansong Xie ◽  
Chadron M. Rose ◽  
Eunbyul Sou ◽  
Hongtao Zeng ◽  
...  
Keyword(s):  
Muscarinic Receptor ◽  
Acinar Cell ◽  
Cell Model ◽  
Ex Vivo ◽  
Receptor Stimulation

scholarly journals Selective BCL-XL Antagonists Eliminate Infected Cells from a Primary Cell Model of HIV Latency but not from Ex Vivo Reservoirs

2020 ◽  
Author(s):  
Yanqin Ren ◽  
Szu Han Huang ◽  
Amanda B. Macedo ◽  
Adam R. Ward ◽  
Winiffer D. Conce Alberto ◽  
...  
Keyword(s):  
T Cells ◽  
Cell Model ◽  
Ex Vivo ◽  
Family Members ◽  
Primary Cell ◽  
Bryostatin 1 ◽  
Viral Reservoirs ◽  
Hiv Persistence ◽  
Hiv Dna ◽  
Infected Cells

AbstractHIV persists, despite antiviral immune responses and effective antiretroviral therapy, in viral reservoirs that seed rebound viremia if therapy is interrupted. Previously, we showed that the BCL-2 protein contributes to HIV persistence by conferring a survival advantage to reservoir-harboring cells. Here, we demonstrate that many of the BCL-2 family members are overexpressed in HIV-infected CD4+ T-cells, indicating increased tension between pro-apoptotic and pro-survival family members – as well as raising the possibility that the inhibition of pro-survival members may disproportionately affect the survival of HIV-infected cells. Based on these results, we chose to further study BCL2L1 (encoding the protein BCL-XL), due to its consistent overexpression and the availability of selective antagonists. Infection of primary CD4+ T-cells with either a clinical isolate, a CCR5-tropic strain, or a CXCR4-tropic strain of HIV resulted in increased BCL-XL protein expression; and treatment with two selective BCL-XL antagonists, A-1155463 and A-1551852, led to disproportionate cell death compared to uninfected CD4+ T-cells. In a primary cell model of latency, both BCL-XL antagonists drove significant reductions in total HIV DNA and in infectious cell frequencies both alone and in combination with the latency reversing agent bryostatin-1, with little off-target cytotoxicity. However, these antagonists, with or without bryostatin-1, or in combination with the highly potent latency reversing agent combination PMA + ionomycin, failed to reduce total HIV DNA and infectious reservoirs in ex vivo CD4+ T-cells from ART-suppressed donors. Our results add to growing evidence that bonafide reservoir-harboring cells are resistant to multiple “kick and kill” modalities - relative to latency models - and uncover BCL-XL antagonists as a facile approach to probing mechanistic underpinnings. We also interpret our results as encouraging of further exploration of BCL-XL antagonists for cure, where combination approaches may unlock the ability to eliminate ex vivo reservoirs.

scholarly journals Detecting spiral wave tips using deep learning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Henning Lilienkamp ◽  
Thomas Lilienkamp
Keyword(s):  
Deep Neural Networks ◽  
Cell Model ◽  
Ex Vivo ◽  
Spiral Wave ◽  
Specific Situation ◽  
Cell Models ◽  
Noise Levels ◽  
Life Threatening ◽  
Spatio Temporal ◽  
Influence Of Noise

AbstractThe chaotic spatio-temporal electrical activity during life-threatening cardiac arrhythmias like ventricular fibrillation is governed by the dynamics of vortex-like spiral or scroll waves. The organizing centers of these waves are called wave tips (2D) or filaments (3D) and they play a key role in understanding and controlling the complex and chaotic electrical dynamics. Therefore, in many experimental and numerical setups it is required to detect the tips of the observed spiral waves. Most of the currently used methods significantly suffer from the influence of noise and are often adjusted to a specific situation (e.g. a specific numerical cardiac cell model). In this study, we use a specific type of deep neural networks (UNet), for detecting spiral wave tips and show that this approach is robust against the influence of intermediate noise levels. Furthermore, we demonstrate that if the UNet is trained with a pool of numerical cell models, spiral wave tips in unknown cell models can also be detected reliably, suggesting that the UNet can in some sense learn the concept of spiral wave tips in a general way, and thus could also be used in experimental situations in the future (ex-vivo, cell-culture or optogenetic experiments).

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