Propagation of Arthropod-Borne Rickettsia spp. in Two Mosquito Cell Lines

ABSTRACT Rickettsiae are obligate intracellular alphaproteobacteria that include pathogenic species in the spotted fever, typhus, and transitional groups. The development of a standardized cell line in which diverse rickettsiae can be grown and compared would be highly advantageous to investigate the differences among and between pathogenic and nonpathogenic species of rickettsiae. Although several rickettsial species have been grown in tick cells, tick cells are more difficult to maintain and they grow more slowly than insect cells. Rickettsia-permissive arthropod cell lines that can be passaged rapidly are highly desirable for studies on arthropod-Rickettsia interactions. We used two cell lines (Aedes albopictus cell line Aa23 and Anopheles gambiae cell line Sua5B) that have not been used previously for the purpose of rickettsial propagation. We optimized the culture conditions to propagate one transitional-group rickettsial species (Rickettsia felis) and two spotted-fever-group rickettsial species (R. montanensis and R. peacockii) in each cell line. Both cell lines allowed the stable propagation of rickettsiae by weekly passaging regimens. Stable infections were confirmed by PCR, restriction digestion of rompA, sequencing, and the direct observation of bacteria by fluorescence in situ hybridization. These cell lines not only supported rickettsial growth but were also permissive toward the most fastidious species of the three, R. peacockii. The permissive nature of these cell lines suggests that they may potentially be used to isolate novel rickettsiae or other intracellular bacteria. Our results have important implications for the in vitro maintenance of uncultured rickettsiae, as well as providing insights into Rickettsia-arthropod interactions.

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Rickettsia felis from Cat Fleas: Isolation and Culture in a Tick-Derived Cell Line

Applied and Environmental Microbiology ◽

10.1128/aem.00532-06 ◽

2006 ◽

Vol 72 (8) ◽

pp. 5589-5595 ◽

Cited By ~ 57

Author(s):

Walairat Pornwiroon ◽

Susan S. Pourciau ◽

Lane D. Foil ◽

Kevin R. Macaluso

Keyword(s):

Cell Line ◽

Citrate Synthase ◽

Ixodes Scapularis ◽

Spotted Fever ◽

Protein A ◽

Etiologic Agent ◽

Host Cells ◽

Spotted Fever Group ◽

Rickettsia Felis ◽

Cat Fleas

ABSTRACT Rickettsia felis, the etiologic agent of spotted fever, is maintained in cat fleas by vertical transmission and resembles other tick-borne spotted fever group rickettsiae. In the present study, we utilized an Ixodes scapularis-derived tick cell line, ISE6, to achieve isolation and propagation of R. felis. A cytopathic effect of increased vacuolization was commonly observed in R. felis-infected cells, while lysis of host cells was not evident despite large numbers of rickettsiae. Electron microscopy identified rickettsia-like organisms in ISE6 cells, and sequence analyses of portions of the citrate synthase (gltA), 16S rRNA, Rickettsia genus-specific 17-kDa antigen, and spotted fever group-specific outer membrane protein A (ompA) genes and, notably, R. felis conjugative plasmids indicate that this cultivatable strain (LSU) was R. felis. Establishment of R. felis (LSU) in a tick-derived cell line provides an alternative and promising system for the expansion of studies investigating the interactions between R. felis and arthropod hosts.

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ANTITUMOR EFFECT OF SUNITINIB AND BORTEZOMIB ON BREAST CANCER CELL LINE IN VITRO

Problems in oncology ◽

10.37469/0507-3758-2017-63-1-141-145 ◽

2017 ◽

Vol 63 (1) ◽

pp. 141-145

Author(s):

Yuliya Khochenkova ◽

Eliso Solomko ◽

Oksana Ryabaya ◽

Yevgeniya Stepanova ◽

Dmitriy Khochenkov

Keyword(s):

Breast Cancer ◽

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Antitumor Effect ◽

Cancer Cell Lines ◽

Breast Cancer Cell Lines ◽

Actual Problem

The discovery for effective combinations of anticancer drugs for treatment for breast cancer is the actual problem in the experimental chemotherapy. In this paper we conducted a study of antitumor effect of the combination of sunitinib and bortezomib against MDA-MB-231 and SKBR-3 breast cancer cell lines in vitro. We found that bortezomib in non-toxic concentrations can potentiate the antitumor activity of sunitinib. MDA-MB-231 cell line has showed great sensitivity to the combination of bortezomib and sunitinib in vitro. Bortezomib and sunitinib caused reduced expression of receptor tyrosine kinases VEGFR1, VEGFR2, PDGFRa, PDGFRß and c-Kit on HER2- and HER2+ breast cancer cell lines

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Design, Synthesis, Molecular Docking, and Anticancer Evaluation of Pyrazole Linked Pyrazoline Derivatives with Carbothioamide Tail as EGFR Kinase Inhibitors

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200727093613 ◽

2020 ◽

Vol 21 (1) ◽

pp. 42-60

Author(s):

Farah Nawaz ◽

Ozair Alam ◽

Ahmad Perwez ◽

Moshahid A. Rizvi ◽

Mohd. Javed Naim ◽

...

Keyword(s):

Molecular Docking ◽

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Line ◽

Cancer Cell Lines ◽

Kinase Assay ◽

Cervix Uteri ◽

Egfr Kinase

Background: The Epidermal Growth Factor Receptor (known as EGFR) induces cell differentiation and proliferation upon activation through the binding of its ligands. Since EGFR is thought to be involved in the development of cancer, the identification of new target inhibitors is the most viable approach, which recently gained momentum as a potential anticancer therapy. Objective: To assess various pyrazole linked pyrazoline derivatives with carbothioamide for EGFR kinase inhibitory as well as anti-proliferative activity against human cancer cell lines viz. A549 (non-small cell lung tumor), MCF-7 (breast cancer cell line), SiHa (cancerous tissues of the cervix uteri), and HCT-116 (colon cancer cell line). Methods: In vitro EGFR kinase assay, in vitro MTT assay, Lactate dehydrogenase release, nuclear staining (DAPI), and flow cytometry cell analysis. Results: Compounds 6h and 6j inhibited EGFR kinase at concentrations of 1.66μM and 1.9μM, respectively. Furthermore, compounds 6h and 6j showed the most potent anti-proliferative results against the A549 KRAS mutation cell line (IC50 = 9.3 & 10.2μM). Through DAPI staining and phase contrast microscopy, it was established that compounds 6h and 6j also induced apoptotic activity in A549 cells. This activity was further confirmed by FACS using Annexin-V-FITC and Propidium Iodide (PI) labeling. Molecular docking studies performed on 6h and 6j suggested that the compounds can bind to the hinge region of ATP binding site of EGFR tyrosine kinase in a similar pose as that of the standard drug gefitinib. Conclusion: The potential anticancer activity of compounds 6h and 6j was confirmed and need further exploration in cancer cell lines of different tissue origin and signaling pathways, as well as in animal models of cancer development.

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Survival and Function of Fibroblasts Stored as Stock Cultures at a Non-freezing Temperature

Alternatives to Laboratory Animals ◽

10.1177/026119299302100215 ◽

1993 ◽

Vol 21 (2) ◽

pp. 206-209

Author(s):

Anders H. G. Andrén ◽

Anders P. Wieslander

Keyword(s):

Cell Growth ◽

Cell Line ◽

Cell Lines ◽

Fibroblast Cell ◽

Freezing Temperature ◽

Mouse Fibroblast ◽

Toxic Response ◽

Normal Manner ◽

And Function

Cytotoxicity, measured as inhibition of cell growth of cultured cell lines, is a widely used method for testing the safety of biomaterials and chemicals. One major technical disadvantage with this method is the continuous routine maintenance of the cell lines. We decided to investigate the possibility of storing stock cultures of fibroblasts (L-929) in an ordinary refrigerator as a means of reducing the routine workload. Stock cultures of the mouse fibroblast cell line L-929 were prepared in plastic vials with Eagle's minimum essential medium. The vials were stored in a refrigerator at 4–10°C for periods of 7–31 days. The condition of the cells after storage was determined as cell viability, cell growth and the toxic response to acrylamide, measured as cell growth inhibition. We found that the L-929 cell line can be stored for 2–3, weeks with a viabilty > 90% and a cell growth of about 95%, compared to L-929 cells grown and subcultured in the normal manner. The results also show that the toxic response to acrylamide, using refrigerator stored L-929 cells, corresponds to that of control L-929 cells. We concluded that it is possible to store L-929 cells in a refrigerator for periods of up to 3 weeks and still use the cells for in vitro cytotoxic assays.

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Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick—Pathogen Interactions

Pathogens ◽

10.3390/pathogens10010070 ◽

2021 ◽

Vol 10 (1) ◽

pp. 70

Author(s):

Lourdes Mateos-Hernández ◽

Natália Pipová ◽

Eléonore Allain ◽

Céline Henry ◽

Clotilde Rouxel ◽

...

Keyword(s):

Cell Line ◽

Peripheral Nerves ◽

Ixodes Scapularis ◽

Embryonic Cell ◽

Cell Subpopulation ◽

In Vivo Experiments

Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like Ixodes scapularis embryonic cell line, ISE6, is an effective tool frequently used for examining tick–pathogen interactions. We detected 37 neuropeptide transcripts in the I. scapularis ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, Anaplasma phagocytophilum. The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in A. phagocytophilum-infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of I. scapularis synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete Ixodes neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick–pathogen interactions.

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Trace Amine-Associated Receptor 1 Trafficking to Cilia of Thyroid Epithelial Cells

Cells ◽

10.3390/cells10061518 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1518

Author(s):

Maria Qatato ◽

Vaishnavi Venugopalan ◽

Alaa Al-Hashimi ◽

Maren Rehders ◽

Aaron D. Valentine ◽

...

Keyword(s):

Cell Surface ◽

Cell Lines ◽

Human Thyroid ◽

Apical Plasma Membrane ◽

Thyroid Cell ◽

Thyroid Cells ◽

Trace Amine ◽

Rat Thyroid

Trace amine-associated receptor 1 (rodent Taar1/human TAAR1) is a G protein-coupled receptor that is mainly recognized for its functions in neuromodulation. Previous in vitro studies suggested that Taar1 may signal from intracellular compartments. However, we have shown Taar1 to localize apically and on ciliary extensions in rodent thyrocytes, suggesting that at least in the thyroid, Taar1 may signal from the cilia at the apical plasma membrane domain of thyrocytes in situ, where it is exposed to the content of the follicle lumen containing putative Taar1 ligands. This study was designed to explore mouse Taar1 (mTaar1) trafficking, heterologously expressed in human and rat thyroid cell lines in order to establish an in vitro system in which Taar1 signaling from the cell surface can be studied in future. The results showed that chimeric mTaar1-EGFP traffics to the apical cell surface and localizes particularly to spherical structures of polarized thyroid cells, procilia, and primary cilia upon serum-starvation. Moreover, mTaar1-EGFP appears to form high molecular mass forms, possibly homodimers and tetramers, in stably expressing human thyroid cell lines. However, only monomeric mTaar1-EGFP was cell surface biotinylated in polarized human thyrocytes. In polarized rat thyrocytes, mTaar1-EGFP is retained in the endoplasmic reticulum, while cilia were reached by mTaar1-EGFP transiently co-expressed in combination with an HA-tagged construct of the related mTaar5. We conclude that Taar1 trafficking to cilia depends on their integrity. The results further suggest that an in vitro cell model was established that recapitulates Taar1 trafficking in thyrocytes in situ, in principle, and will enable studying Taar1 signaling in future, thus extending our general understanding of its potential significance for thyroid autoregulation.

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Tissue type-specific expression of intermediate filament proteins in a cultured epithelial cell line from bovine mammary gland

The Journal of Cell Biology ◽

10.1083/jcb.96.1.37 ◽

1983 ◽

Vol 96 (1) ◽

pp. 37-50 ◽

Cited By ~ 93

Author(s):

E Schmid ◽

DL Schiller ◽

C Grund ◽

J Stadler ◽

WW Franke

Keyword(s):

Mammary Gland ◽

Epithelial Cell ◽

Cell Line ◽

Cell Lines ◽

Intermediate Filament ◽

Striking Difference ◽

Intermediate Filament Proteins ◽

Cell Clones ◽

Vimentin Filaments

Different clonal cell lines have been isolated from cultures of mammary gland epithelium of lactating cow's udder and have been grown in culture media containing high concentrations of hydrocortisone, insulin, and prolactin. These cell (BMGE+H), which grow in monolayers of typical epithelial appearance, are not tightly packed, but leave intercellular spaces spanned by desmosomal bridges. The cells contain extended arrays of cytokeratin fibrils, arranged in bundles attached to desmosomes. Gel electophoresis show that they synthesize cytokeratins similar, if not identical, to those found in bovine epidermis and udder, including two large (mol wt 58,500 and 59,000) and basic (pH range: 7-8) and two small (mol wt 45,500 and 50,000) and acidic (pH 5.32 and 5.36) components that also occur in phosphorylated forms. Two further cytokeratins of mol wts 44,000 (approximately pH 5.7) and 53,000 (pH 6.3) are detected as minor cytokeratins in some cell clones. BMGE+H cells do not produce vimentin filaments as determined by immunofluorescence microscopy and gel electrophoresis. By contrast, BMGE-H cells, which have emerged from the same original culture but have been grown without hormones added, are not only morphologically different, but also contain vimentin filaments and a different set of cytokeratins, the most striking difference being the absence of the two acidic cytokeratins of mol wt 50,000 and 45,500. Cells of the BMGE+H line are characterized by an unusual epithelial morphology and represent the first example of a nonmalignant permanent cell line in vitro that produces cytokeratin but not vimentin filaments. The results show that (a) tissue-specific patterns of intermediate filament expression can be maintained in permanent epithelial cell lines in culture, at least under certain growth conditions; (b) loss of expression of relatively large, basic cytokeratins is not an inevitable consequence of growth of epithelial cells in vitro. Our results further show that, during culturing, different cell clones with different cytoskeletal composition can emerge from the same cell population and suggest that the presence of certain hormones may have an influence on the expression of intermediate filament proteins.

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Synthesis and Experimental Validation of New Designed Heterocyclic Compounds with Antiproliferative Activity versus Breast Cancer Cell Lines MCF-7 and MDA-MB-231

Journal of Chemistry ◽

10.1155/2017/9729284 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 2

Author(s):

Vincenza Barresi ◽

Carmela Bonaccorso ◽

Domenico A. Cristaldi ◽

Maria N. Modica ◽

Nicolò Musso ◽

...

Keyword(s):

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Anticancer Agents ◽

Cancer Cell Line ◽

Breast Cancer Cell Lines ◽

Design And Synthesis ◽

Human Breast Cell ◽

Mcf 7

Recent drug discovery efforts are highly focused towards identification, design, and synthesis of small molecules as anticancer agents. With this aim, we recently designed and synthesized novel compounds with high efficacy and specificity for the treatment of breast tumors. Based on the obtained results, we constructed a Volsurf+ (VS+) model using a dataset of 59 compounds able to predict the in vitro antitumor activity against MCF-7 cancer cell line for new derivatives. In the present paper, in order to further verify the robustness of this model, we report the results of the projection of more than 150 known molecules and 9 newly synthesized compounds. We predict their activity versus MCF-7 cell line and experimentally verify the in silico results for some promising chosen molecules in two human breast cell lines, MCF-7 and MDA-MB-231.

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Expression of the SMP antigen by oligodendrocytes in the developing avian central nervous system

Development ◽

10.1242/dev.107.4.825 ◽

1989 ◽

Vol 107 (4) ◽

pp. 825-833

Author(s):

P. Cameron-Curry ◽

C. Dulac ◽

N.M. Le Douarin

Keyword(s):

Monoclonal Antibody ◽

Schwann Cell ◽

Basic Protein ◽

Culture Conditions ◽

Cell Marker ◽

Cellular Expression ◽

Early Expression ◽

Different Parts

Expression of the avian antigen SMP (Schwann cell Myelin Protein, Mr 75-80000), first characterized in the PNS with a monoclonal antibody as an early and strictly specific Schwann cell marker, was further studied in the CNS. Comparing SMP immunoreactive areas in the different parts of the CNS with those expressing the Myelin Basic Protein (MBP), we showed a strict colocalisation of both phenotypes. In vitro, MBP+ oligodendrocytes express the surface antigen SMP as well. SMP cellular expression was followed in situ and in culture using nervous tissues from embryos at different stages. We were thus able to detect an early expression of this marker by oligodendroblasts before the first appearance of MBP immunoreactivity. We have also identified a subpopulation of SMP+/MBP- and SMP+/GC- cells, which persists under our culture conditions as precursors remaining in an immature state.

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Vitamin D as Radiosensitizer: A Review in Cell Line -

Journal of Pharmacy and Nutrition Sciences ◽

10.29169/1927-5951.2020.10.06.1 ◽

2020 ◽

Vol 10 (6) ◽

pp. 315-324

Author(s):

Fahmi Radityamurti ◽

Fauzan Herdian ◽

Tiara Bunga Mayang Permata ◽

Handoko Handoko ◽

Henry Kodrat ◽

...

Keyword(s):

Vitamin D ◽

Cell Differentiation ◽

Cell Line ◽

Cell Lines ◽

Anticancer Agent ◽

Medical Literature ◽

In Vitro Studies ◽

Anti Cancer

Introduction: Vitamin D has been shown to have anti-cancer properties such as antioxidants, anti-proliferative, and cell differentiation. The property of vitamin D as an anticancer agent triggers researchers to find out whether vitamin D is useful as a radiosensitizer. Multiple studies have been carried out on cell lines in various types of cancer, but the benefits of vitamin D as a radiosensitizer still controversial. This paperwork aims to investigate the utilization of Vitamin D3 (Calcitriol) as radiosensitizer in various cell line through literature review.Methods: A systematic search of available medical literature databases was performed on in-vitro studies with Vitamin D as a radiosensitizer in all types of cell lines. A total of 11 in-vitro studies were evaluated.Results: Nine studies in this review showed a significant effect of Vitamin D as a radiosensitizer agent by promoting cytotoxic autophagy, increasing apoptosis, inhibiting of cell survival and proliferation, promoting gene in ReIB inhibition, inducing senescene and necrosis. The two remaining studies showed no significant effect in the radiosensitizing mechanism of Vitamin D due to lack of evidence in-vitro settings.Conclusion: Vitamin D have anticancer property and can be used as a radiosensitizer by imploring various mechanism pathways in various cell lines. Further research especially in-vivo settings need to be evaluated.

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