You are interested in our poster presentations or product flyers and brochures? We take pride in our competent service and swift response. Please do not hesitate to contact us for further information. We also very much welcome your feedback and comments. It is of great challenge to predict human brain penetration for substrates of multidrug resistance protein 1 MDR1 and breast cancer resistance protein BCRP , 2 major efflux transporters at blood-brain barrier. In this study, 2 widely used human MDR1 cell lines from Borst and National Institutes of Health laboratories were evaluated using rodent brain penetration data, and the study suggested that the MDR1 expressed in Madin-Darby canine kidney MDCK cell line from National Institutes of Health laboratory predicted brain penetration better, particularly for compounds with a high passive permeability.
COM0 1. COM6 1. In addition to cloned MDCK cells the indicator cell lines used to evaluate the impact of producing LAIV in cells on host range and replication included two human cell lines: human lung carcinoma A cells and human muco-epidermoid bronchiolar carcinoma NCI H cells. Results from this study demonstrate remarkable similarity between influenza viruses representing the current commercial egg produced and developmental MDCK cell produced vaccine production platforms. MedImmune's high yielding cloned MDCK cells used for the cell culture based vaccine production were highly permissive to both egg and cell produced ca attenuated influenza viruses.
Irrespective of the indicator cell line used the replication properties were similar between egg and the cell produced. Si-Ni-San SNS possesses extensive therapeutic effects, however, the extent to which main components are absorbed and the mechanisms involved are controversial. In this study, MDCK cell model was used to determine the permeability characteristics and interaction between the major components of Si-Ni-San, including saikosaponin a, paeoniflorin, naringin and glycyrrhizic acid.
The transport of the major components was concentration-dependent in both directions. Additionally, the efflux of paeoniflorin and naringin were apparently reduced in the presence of P-gp inhibitor verapamil. The transport of glycyrrhizic acid was clearly inhibited by the inhibitors of MRP2, indicating that MRP2 may be involved in the transport of glycyrrhizic acid.
However, the results indicated that saikosaponin a was absorbed mainly by passive diffusion. Furthermore, the combined incubation of four major components had a powerful sorbefacient effect than a single drug used alone which may be regulated by tight junctions. Taken together, our study provides useful information for pharmacological applications of Si-Ni-San and offers new insights into this ancient decoction for further researches, especially in drug synergism.
Epithelial to mesenchymal transition EMT occurs during embryogenesis or under pathological conditions such as hypoxia, injury, chronic inflammation, or tissue fibrosis. These results suggest a putative role of purine receptors as target for anti-fibrotic agents.
Vaccination remains the principal measure to reduce morbidity and mortality from such pandemics. The availability and surging demand for pandemic vaccines needs to be addressed in the preparedness plans. This study presents an improved high-yield manufacturing process for the inactivated influenza H5N1 vaccines using Madin-Darby canine kidney MDCK cells grown in a serum-free SF medium microcarrier cell culture system. The selected SF medium was further evaluated in various bioreactor culture systems for process scale-up evaluation.
No significant difference was found in the cell growth in different sizes of bioreactors studied. In the 7. A mouse immunogenicity study showed that the formalin-inactivated purified SF vaccine candidate formulated with alum adjuvant could induce protective level of virus neutralization titers similar to those obtained from the SC medium. This study provides useful information to manufacturers that are planning to use SF medium for cell -based influenza vaccine production.
Full Text Available Abstract Background In cell culture-based influenza vaccine production the monitoring of virus titres and cell physiology during infection is of great importance for process characterisation and optimisation. While conventional virus quantification methods give only virus titres in the culture broth, data obtained by fluorescence labelling of intracellular virus proteins provide additional information on infection dynamics.
Flow cytometry represents a valuable tool to investigate the influences of cultivation conditions and process variations on virus replication and virus yields. Results In this study, fluorescein-labelled monoclonal antibodies against influenza A virus matrix protein 1 and nucleoprotein were used for monitoring the infection status of adherent Madin-Darby canine kidney cells from bioreactor samples.
Monoclonal antibody binding was shown for influenza A virus strains of different subtypes H1N1, H1N2, H3N8 and host specificity human, equine, swine. At high multiplicity of infection in a bioreactor, the onset of viral protein accumulation in adherent cells on microcarriers was detected at about 2 to 4 h post infection by flow cytometry. In contrast, a significant increase in titre by hemagglutination assay was detected at the earliest 4 to 6 h post infection.
Conclusion It is shown that flow cytometry is a sensitive and robust method for the monitoring of viral infection in fixed cells from bioreactor samples. Therefore, it is a valuable addition to other detection methods of influenza virus infection such as immunotitration and RNA hybridisation.
Thousands of individual cells are measured per sample. Thus, the presented method is believed to be quite independent of the concentration of infected cells multiplicity of infection and total cell concentration in bioreactors. This allows to perform detailed studies on factors relevant for optimization of virus yields in cell cultures. The method could also be used for process. The human colon adenocarcinoma Caco-2 and Madin-Darby canine kidney epithelial cell lines provide in vitro tools to assess a drug's permeability and transporter interactions during discovery and development.
The cells , when cultured on semiporous filters, form confluent monolayers that model the intestinal epithelial barrier for permeability, transporter and drug-interaction assays. The applications of these assays in pharmaceutical research include qualitative prediction and ranking of absorption, determining mechanism s of permeability, formulation effects on drug permeability, and the potential for transporter-mediated drug-drug interactions.
This review focuses on recent examples of Caco-2 and Madin-Darby canine kidney cells assays for drug permeability including transfected and knock-down cells , miniaturization and automation, and assay combinations to better understand and predict intestinal drug absorption. Annual influenza epidemics continue to have a considerable impact in both developed and developing countries.
Vaccination remains the principal measure to prevent seasonal influenza and reduce associated morbidity and mortality. The WHO recommends using established mammalian cell culture lines as an alternative to egg-based substrates in the manufacture of influenza vaccine. This review examines the advantages and disadvantages of cell culture-based technology for influenza vaccine production, compares immunogenicity and safety data for Optaflu with that of currently marketed conventional egg-based influenza vaccines, and considers the prospects for wider use of cell culture-based influenza vaccines.
Full Text Available Cyst enlargement in polycystic kidney disease PKD involves cAMP-activated proliferation of cyst-lining epithelial cells and transepithelial fluid secretion into the cyst lumen via cystic fibrosis transmembrane conductance regulator CFTR chloride channel. This study aimed to investigate an inhibitory effect and detailed mechanisms of steviol and its derivatives on cyst growth using a cyst model in Madin-Darby canine kidney MDCK cells. Among 4 steviol-related compounds tested, steviol was found to be the most potent at inhibiting MDCK cyst growth.
Steviol inhibition of cyst growth was dose-dependent; steviol microM reversibly inhibited cyst formation and cyst growth by However, steviol acutely inhibited forskolin-stimulated apical chloride current in MDCK epithelia, measured with the Ussing chamber technique, in a dose-dependent manner.
Prolonged treatment 24 h with steviol microM also strongly inhibited forskolin-stimulated apical chloride current, in part by reducing CFTR protein expression in MDCK cells. Immunofluorescence studies demonstrated that prolonged treatment 24 h with steviol microM markedly reduced CFTR expression at the plasma membrane. Steviol and related compounds therefore represent drug candidates for treatment of polycystic kidney disease.
Dekker, E. The localization of. Dynamic bio-adhesion of polymer nanoparticles on MDCK epithelial cells and its impact on bio-membranes, endocytosis and paracytosis. Nowadays, concern about the use of nanotechnology for biomedical application is unprecedentedly increasing. In fact, nanosystems applied for various potential clinical uses always have to cross the primary biological barrier consisting of epithelial cells.
However, little is really known currently in terms of the influence of the dynamic bio-adhesion of nanosystems on bio-membranes as well as on endocytosis and transcytosis. Firstly, the adhesion of PNs on cell membranes was found to be time-dependent with a shift of both location and dispersion pattern, from the lateral adhesion of mainly mono-dispersed PNs initially to the apical coverage of the PN aggregate later.
Then, it was interesting to observe in this study that the dynamic bio-adhesion of PNs only affected their endocytosis but not their transcytosis. It was important to find that the endocytosis of PNs was not a constant process. A GM1 dependent CDE caveolae dependent endocytosis pathway was dominant in the preliminary stage, followed by the co-existence of a CME clathrin-mediated endocytosis pathway for the PN aggregate at a later stage, in accordance with the adhesion features of PNs, suggesting the modification of PN adhesion patterns on the endocytosis pathways.
Next, the PN adhesion was noticed to affect the structure of cell junctions, via altering the extra- and intra-cellular calcium levels, leading to the enhanced paracellular transport of small molecules, but not favorably enough for the obviously increased passing of PNs themselves.
Finally, FRAP and other techniques all demonstrated the obvious impact of PN adhesion on the membrane confirmation, independent of the adhesion location and time, which might lower the threshold for the internalization of PNs, even their aggregates. Generally, these findings confirm that the transport pathway mechanism of PNs through epithelial cells is rather.
Alkaloids have been demonstrated to be the predominant pharmacological active components of URCU. The samples were analyzed by high-performance liquid chromatography, and the apparent permeability coefficients Papp were calculated. The time- and concentration-dependency experiments indicated that the main mechanism for 2, 4, 5 and 6 through BBB was passive diffusion.
The efflux mechanism involved in the transports of compounds 1 and 3 could be reduced significantly by verapamil, and molecular docking screening also showed that 1 and 3 had strong bindings to P-glycoprotein. This study provides useful information for predicting the BBB permeability for 1—6, as well as better understanding of their central nervous system pharmacological activities.
So far, the content and accumulation of ATP in isolated endoplasmic reticulum ER are little understood. First, we confirmed using electron microscopic and Western blotting techniques that the samples extracted from MDCK cells are endoplasmic reticulum ER.
The amounts of ATP in the extracted ER were measured from the filtrate after a spinning down of ultrafiltration spin column packed with ER. Cyclooxygenase expression in canine platelets and Madin-Darby canine kidney cells. Canine platelets and MDCK cells. Following northern blot analysis, canine platelets were found to express only the 2. Canine MDCK cells expressed the 4. Cloning and sequencing of the canine gene will be required to fully characterize homologous regions.
Because of the importance of COX in the inflammatory process and as a potential target of currently available nonsteroidal anti-inflammatory drugs NSAID , a better understanding of canine COX may improve our ability to use NSAID appropriately, achieve efficacy, and avoid potential adverse drug effects in dogs. The fruits of Euodia rutaecarpa Euodiae Fructus, EF , the widely used traditional Chinese medicine, have various central nervous system effects.
The transport samples were analyzed by high performance liquid chromatography and the apparent permeability coefficients P app were calculated. Cell sources for BBB models include primary brain endothelial cells or immortalized brain endothelial cell lines. Despite their well-known differences, epithelial cell lines are also used as surrogate models for testing neuropharmaceuticals.
The aim of the present study was to compare the expression of selected BBB related genes including tight junction proteins, solute carriers SLC, ABC transporters, metabolic enzymes and to describe the paracellular properties of nine different culture models. To establish a primary BBB model rat brain capillary endothelial cells were co-cultured with rat pericytes and astrocytes EPA.
To test transporter functionality, the permeability of 12 molecules, glucopyranose, valproate, baclofen, gabapentin, probenecid, salicylate, rosuvastatin, pravastatin, atorvastatin, tacrine, donepezil, was also measured in the EPA and epithelial models. Among the junctional protein genes, the expression level of occludin was high in all models except the GP8 and RBE4 cells , and each model expressed a unique claudin pattern.
Full Text Available P-glycoprotein, a human multidrug resistance transporter, has been extensively studied due to its importance to human health and disease. In order to understand transport kinetics via P-gp, confluent cell monolayers overexpressing P-gp are widely used.
The purpose of this study is to obtain the mass action elementary rate constants for P-gp's transport and to functionally characterize members of P-gp's network, i. Transport of a range of concentrations of amprenavir, loperamide, quinidine and digoxin across the confluent monolayer of cells was measured in both directions, apical to basolateral and basolateral to apical.
We developed a global optimization algorithm using the Particle Swarm method that can simultaneously fit all datasets to yield accurate and exhaustive fits of these elementary rate constants. The statistical sensitivity of the fitted values was determined by using 24 identical replicate fits, yielding simple averages and standard deviations for all of the kinetic parameters, including the efflux active P-gp surface density.
Digoxin required additional basolateral and apical transporters, while loperamide required just a basolateral tranporter. The data were better fit by assuming bidirectional transporters, rather than active importers, suggesting that they are not MRP or active OATP transporters. The P-gp efflux rate constants for quinidine and digoxin were about 3-fold smaller than reported ATP hydrolysis rate constants from P-gp proteoliposomes.
The fitted values of the elementary rate constants for these P-gp substrates support the hypotheses that the selective pressures on P-gp are to maintain a broad substrate range and to keep xenobiotics out of the cytosol, but not out of the. The potassium channel K V 7. In addition, K V 7.
Mutations in the kcnq1 gene can lead to long QT syndrome To learn more about the basic mechanisms that regulate K V 7. Molecular evidence and functional expression of multidrug resistance associated protein MRP in rabbit corneal epithelial cells. Multidrug resistance associated protein MRP is a major family of efflux transporters involved in drug efflux leading to drug resistance. The objective of this study was to explore physical barriers for ocular drug absorption and to verify if the role of efflux transporters.
Membrane fraction of rCEC was used for western blot analysis. For the first time we have demonstrated high expression of MRP-2 in rabbit corneal epithelium and its functional activity causing drug efflux. Cooperation between epithelial cells demonstrated by potassium transfer. We now report the extension of this assay procedure to cultured epithelial cells. The degree of enhancement, expressed as index of cooperation, depended on the numbers of cells in the cultures, their opportunity for cell-to-cell contact, and above a certain permissive level the concentration of ouabain.
Autoradiography of cells incubated with [3H]leucine demonstrated that MDCK cells in ouabain-treated mixed cultures were able to synthesize proteins only when physically adjacent to HaK cells. The transmission of labeled nucleosides among the cells provides independent evidence of the phenomenon of cooperation, probably mediated by gap junctions. This system offers promise for investigation of stimuli modulating junctional communication. Tetraethylammonium block of water flux in Aquaporin-1 channels expressed in kidney thin limbs of Henle's loop and a kidney-derived cell line.
Full Text Available Abstract Background Aquaporin-1 AQP1 channels are constitutively active water channels that allow rapid transmembrane osmotic water flux, and also serve as cyclic-GMP-gated ion channels. Tetraethylammonium chloride TEA; 0. We also demonstrate that TEA does not inhibit the cGMP-dependent ionic conductance of AQP1 expressed in oocytes, supporting the idea that water and ion fluxes involve pharmacologically distinct pathways in the AQP1 tetrameric complex.
Results TEA blocked water permeability of AQP1 channels in kidney and kidney-derived cells , demonstrating this effect is not limited to the oocyte expression system. Conclusions TEA selectively inhibits osmotic water permeability through native and heterologously expressed AQP1 channels. The pathways for water and ions in AQP1 differ in pharmacological sensitivity to TEA, and are consistent with the idea of independent solute pathways within the channel structure.
Polarized expression of the GFP-tagged rat V 1a vasopressin receptor. Therefore, the GFP-tagged V 1a receptor retains all the sorting signals of the wild-type receptor and offers an excellent system to elucidate the mechanisms of cell trafficking of V 1a receptors. The solubility enhancing effects of various excipients, including their compatibility with in vitro permeability P app systems, was investigated using drugs representative of Biopharmaceutics Classification System BCS classes I-IV.
Turbidimetric solubility determination using nephelometry and transport experiments using MDCK Strain I cell monolayers were employed. The highest usable concentration of each excipient [dimethyl sulfoxide DMSO , ethanol, hydroxypropyl-beta-cyclodextrin HPCD , and sodium taurocholate] was determined by monitoring apical AP to basolateral BL [14C]mannitol apparent permeability P app and the transepithelial electrical resistance TEER in transport experiments done at pH 6.
Additionally, a novel in vitro system aimed at more accurately simulating in vivo conditions, i. We demonstrate that K 7. Our data provide a possible explanation To construct recombinant adenovirus containing canine interferon-gamma cIFN-gamma gene and to investigate its antiviral activity against canine parvovirus in Madin-Darby canine kidney cells MDCK. To analyze its anti-canine parvovirus activity, the MDCK cells were pre-infected by Ad-cIFN-gamma recombinant adenovirus, and then infected by canine parvovirus.
The antiviral activity of the Ad-cIFN-gamma recombinant adenovirus against parvovirus was analyzed. The recombinant adenovirus containing cIFN-gamma gene was constructed by the ligation method. The Ad-cIFN-gamma recombinant adenovirus could significantly inhibit canine parvovirus replication in MDCK cells pre-infected with the recombinant adenovirus. These results indicate that the Ad-cIFN-gamma recombinant adenovirus has the potent antiviral activity against canine parvovirus.
The Ad-cIFN-gamma recombinant adenovirus was successfully constructed by the ligation method and possessed a powerful antiviral activity against canine parvovirus. Renal tissues are subject to metabolic and respiratory acidosis, and acidosis has been shown to acutely activate NHE1 activity in other cell types.
These results suggest that acute acidosis activates NHE1 in mammalian kidney cells and that in MDCK cells this activation occurs through an ERK-dependent pathway affecting phosphorylation of a distinct set of amino acids in the cytosolic regulatory tail of NHE1. In epithelia that have active transport functions, the force for transmembrane flux of an ion is dictated by the electrochemical gradient in which TEP plays an essential role.
This endogenous electric field is implicated to enhance wound healing by guiding cell migration. We thus seek techniques to enhance the TEP, which may increase the wound electric fields and enhance wound healing. SM significantly increased TEP over four fold. Our study for the first time developed an electrical approach to significantly increase the TEP. Rho GTPase expression in human myeloid cells.
Full Text Available Myeloid cells are critical for innate immunity and the initiation of adaptive immunity. Strict regulation of the adhesive and migratory behavior is essential for proper functioning of these cells. Rho GTPases are important regulators of adhesion and migration; however, it is unknown which Rho GTPases are expressed in different myeloid cells. In addition, the Rho GTPase expression profile changes during dendritic cell maturation with Rac1 being upregulated and Rac2 downregulated.
Our data uncover cell type specific modulation of the Rho GTPase expression profile in hematopoietic stem cells and in more differentiated cells of the myeloid lineage. Foxp3 expression in human cancer cells. Full Text Available Abstract Objective Transcription factor forkhead box protein 3 Foxp3 specifically characterizes the thymically derived naturally occurring regulatory T cells Tregs.
Limited evidence indicates that it is also expressed , albeit to a lesser extent, in tissues other than thymus and spleen, while, very recently, it was shown that Foxp3 is expressed by pancreatic carcinoma. This study was scheduled to investigate whether expression of Foxp3 transcripts and mature protein occurs constitutively in various tumor types. Materials and methods Twenty five tumor cell lines of different tissue origins lung cancer, colon cancer, breast cancer, melanoma, erythroid leukemia, acute T- cell leukemia were studied.
Results Foxp3 mRNA as well as Foxp3 protein was detected in all tumor cell lines, albeit in variable levels, not related to the tissue of origin. Conclusion We offer evidence that Foxp3 expression , characterizes tumor cells of various tissue origins. The biological significance of these findings warrants further investigation in the context of tumor immune escape, and especially under the light of current anti-cancer efforts interfering with Foxp3 expression.
Decorin expression in quiescent myogenic cells. Satellite cells are quiescent muscle stem cells that promote postnatal muscle growth and repair. When satellite cells are activated by myotrauma, they proliferate, migrate, differentiate, and ultimately fuse to existing myofibers.
The remainder of these cells do not differentiate, but instead return to quiescence and remain in a quiescent state until activation begins the process again. This ability to maintain their own population is important for skeletal muscle to maintain the capability to repair during postnatal life.
However, the mechanisms by which satellite cells return to quiescence and maintain the quiescent state are still unclear. Here, we demonstrated that decorin mRNA expression was high in cell cultures containing a higher ratio of quiescent satellite cells when satellite cells were stimulated with various concentrations of hepatocyte growth factor. This result suggests that quiescent satellite cells express decorin at a high level compared to activated satellite cells. Furthermore, we examined the expression of decorin in reserve cells , which were undifferentiated myoblasts remaining after induction of differentiation by serum-deprivation.
Decorin mRNA levels in reserve cells were higher than those in differentiated myotubes and growing myoblasts. These results suggest that decorin participates in the quiescence of myogenic cells. AMP-activated protein kinase downregulates Kv7. In this study, the pathway downstream of PKC, which leads to internalization of Kv7. We show by confocal We demonstrate Hakai reduces cell -substratum adhesion and increases epithelial cell invasion. The dynamic regulation of cell-cell adhesions is crucial for developmental processes, including tissue formation, differentiation and motility.
Adherens junctions are important components of the junctional complex between cells and are necessary for maintaining cell homeostasis and normal tissue architecture. E-cadherin is the prototype and best-characterized protein member of adherens junctions in mammalian epithelial cells.
Regarded as a tumour suppressor, E-cadherin loss is associated with poor prognosis in carcinoma. The E3 ubiquitin-ligase Hakai was the first reported posttranslational regulator of the E-cadherin complex. Hakai specifically targetted E-cadherin for internalization and degradation and thereby lowered epithelial cell-cell contact. Hakai was also implicated in controlling proliferation, and promoted cancer-related gene expression by increasing the binding of RNA-binding protein PSF to RNAs encoding oncogenic proteins.
We sought to investigate the possible implication of Hakai in cell -substratum adhesions and invasion in epithelial cells. Western blot and immunofluoresecence analyses were performed to assess the roles of Paxillin, FAK and Vinculin in cell -substratum adhesion. The role of the proteasome in controlling cell -substratum adhesion was studied using two proteasome inhibitors, lactacystin and MG Here, we present evidence that implicate Hakai in reducing cell -substratum adhesion and increasing epithelial cell invasion, two hallmark features of cancer progression and metastasis.
Paxillin, an important protein component of the cell -matrix adhesion, was completely absent from focal adhesions and focal contacts in Hakai-overexpressing MDCK cells. Full Text Available A large body of work supports the proposal that transplantation of olfactory ensheathing cells OECs into nerve or spinal cord injuries can promote axonal regeneration and remyelination. Yet, some investigators have questioned whether the transplanted OECs associate with axons and form peripheral myelin, or if they recruit endogenous Schwann cells that form myelin.
Olfactory bulbs from transgenic mice expressing the enhanced green fluorescent protein eGFP under the control of the cyclic nucleotide 3-phosphodiesterase CNPase promoter were studied. CNPase is expressed in myelin-forming cells throughout their lineage.
Transplantation of OECs into transected peripheral nerve longitudinally associated with the regenerated axons. Thus, while OECs do not normally form myelin on olfactory nerve axons, their expression of CNPase is commensurate with their potential to form myelin when transplanted into injured peripheral nerve.
Changes in ceramide metabolism are essential in Madin-Darby canine kidney cell differentiation. Ceramides Cers and complex sphingolipids with defined acyl chain lengths play important roles in numerous cell processes.
Six Cer synthase CerS isoenzymes CerS are the key enzymes responsible for the production of the diversity of molecular species. In this study, we investigated the changes in sphingolipid metabolism during the differentiation of Madin-Darby canine kidney MDCK cells. We next evaluated the different synthesis pathways with sphingolipid inhibitors and found that cells subjected to hypertonicity in the presence of amitriptyline, an inhibitor of acid sphingomyelinase, showed decreased radiolabeled incorporation in LacCer and cells did not develop a mature apical membrane.
These results suggest that hypertonicity induces the endolysosomal degradation of SM, generating the Cer used as substrate for the synthesis of specific molecular species of glycosphingolipids that are essential for MDCK cell differentiation.
Differential expression of cell adhesion genes. The genes whose expression correlates with poor If the proteins involved in tethering cells to the extracellular matrix are important in conferring drug resistance, it may be possible to improve chemotherapy by designing drugs that target these proteins Epithelial cell polarization involves several kinase signaling cascades that eventually divide the surface membrane into an apical and a basolateral part.
One kinase, which is activated during the polarization process, is phosphoinositide 3-kinase PI3K. In MDCK cells , the basolateral potassium Here, we demonstrate that Kv7. Pharmacological inhibition of SGK1 gave similar results Furthermore, knockdown of the ubiquitin ligase Nedd overruled PI3K inhibition, whereas a Nedd interaction-deficient Kv7.
Transitional cell carcinoma express vitamin D receptors. Recently, vitamin D analogues have shown antineoplastic effect in several diseases. The purpose of the present study was therefore Similarly, also tumor grade appeared to be related to the number of cells expressing the receptor. Normal urothlium also expressed VDR but only with low intensity.
Our study shows that TCC cells possess the VDR receptor which may make them capable to respond to stimulation with vitamin D, but functional Merkel cell carcinoma MCC is a malignant neuroendocrine skin tumor frequently associated with the Merkel cell polyomavirus. Immune checkpoint therapy showed remarkable results, although not all patients are responsive to this therapy. Anti-tropomyosin receptor kinase A TrkA -targeted treatment has shown promising results in several tumor entities.
Thirty-nine of the 55 samples were suitable for further histopathologic examination. Expression of TrkA was explored by immunohistochemical analysis. Expression of TrkA on the tumor cells. Specimens of 39 patients 21 women and 18 men; mean [SD] age, This result may lead to the exploration of new targeted treatment options in MCC, especially for patients who do not respond to anti-programmed cell death protein 1 treatment.
Celastrol inhibits the proliferation of a variety of tumor cells including leukemia, glioma, prostate, and breast cancer; however, the possible role of celastrol in the EMT is unclear. We investigated the effect of celastrol on the EMT. They play a key role in several biological processes.
Their abnormal expression may lead to malignant cell transformation. This led to alterations in their mRNA-target. Dissecting stem cell differentiation using single cell expression profiling. Many assumptions about the way cells behave are based on analyses of populations. However, it is now widely recognized that even apparently pure populations can display a remarkable level of heterogeneity.
This is particularly true in stem cell biology where it hinders our understanding of normal development and the development of strategies for regenerative medicine. Over the past decade technologies facilitating gene expression analysis at the single cell level have become widespread, provi Dental enamel cells express functional SOCE channels. Cell cycle gene expression under clinorotation. Cyclins and cyclin-dependent kinase CDK are main regulators of the cell cycle of eukaryotes.
It's assumes a significant change of their level in cells under microgravity conditions and by other physical factors actions. The clinorotation use enables to determine the influence of gravity on simulated events in the cell during the cell cycle - exit from the state of quiet stage and promotion presynthetic phase G1 and DNA synthesis phase S of the cell cycle.
For the clinorotation effect study on cell proliferation activity is the necessary studies of molecular mechanisms of cell cycle regulation and development of plants under altered gravity condition. The activity of cyclin D, which is responsible for the events of the cell cycle in presynthetic phase can be controlled by the action of endogenous as well as exogenous factors, but clinorotation is one of the factors that influence on genes expression that regulate the cell cycle.
These data can be used as a model for further research of cyclin - CDK complex for study of molecular mechanisms regulation of growth and proliferation. In this investigation we tried to summarize and analyze known literature and own data we obtained relatively the main regulators of the cell cycle in altered gravity condition.
Full Text Available Affine chromatography was used to isolate Lec antibodies from the sera of a healthy female donor with the high titers of these anti- bodies, which were labeled with biotin. The study enrolled 51 patients with primary breast cancer BC. Antigen expression was found by immunohistochemistry and flow cytometry. Metallothionein gene expression in renal cell carcinoma. In general, MT is known to modulate three fundamental processes: 1 the release of gaseous mediators such as hydroxyl radical or nitric oxide, 2 apoptosis and 3 the binding and exchange of heavy metals such as zinc, cadmium or copper.
Previous studies have shown a positive correlation between the expression of MT with invasion, metastasis and poor prognosis in various cancers. Most of the previous studies primarily used immunohistochemistry to analyze localization of MT in renal cell carcinoma RCC. Corresponding adjacent normal renal parenchyma was taken as control. All statistical calculations were performed using SPSS software.
Localization of influenza virus proteins to nuclear dot 10 structures in influenza virus-infected cells. GFP-M1 was incorporated into virions produced by influenza virus infected MDCK cells expressing the fusion protein indicating that the fusion protein is at least partially functional. Immunofluorescence revealed that the nuclear dots represent nuclear dot 10 ND10 structures.
These findings demonstrate a previously unappreciated involvement of influenza virus with ND10, a structure involved in cellular responses to immune cytokines as well as the replication of a rapidly increasing list of viruses. Cholinergic regulation of VIP gene expression in human neuroblastoma cells. Vasoactive intestinal polypeptide, muscarinic receptor, neuroblastoma cell , mRNA, gene expression , peptide processing Many genes are expressed in bursts, which can contribute to cell-to-cell heterogeneity.
It is now possible to measure this heterogeneity with high throughput single cell gene expression assays single cell qPCR and RNA-seq. These experimental approaches generate gene expression distributions which can be used to estimate the kinetic parameters of gene expression bursting, namely the rate that genes turn on, the rate that genes turn off, and the rate of transcription.
We construct a complete Izumi, Hiroto, E-mail: h-izumi med. To assess the function of ZNF expression in the cell cycle, we established two cells with forced expression of ZNF derived from PC3 prostate cancer cell lines. However, the doubling time of cells with forced expression of ZNF was approximately twice as long as its control counterpart cell line.
Analysis following serum starvation and re-seeding showed that PC3 cells were synchronized at G1 in the cell cycle. These data suggested that fluctuations in ZNF expression are required both for gene expression associated with cell cycle and for cell division. Caveolin-1 down-regulation is required for Wnt5a-Frizzled 2 signalling in Ha-RasV12 -induced cell transformation. Cav1 overexpression abrogates the Ha-Ras V12 -driven transformation of MK4 cells ; however, the targeted down-regulation of Cav1 is not sufficient to mimic this transformation.
Nanoparticle tracking analysis showed that Ha-Ras V12 -inducing MK4 cells increased exosome-like microvesicles release compared with their normal counterparts. These data suggest that Cav1-dependent repression of Fzd2 and exosome uptake is potentially relevant to its antitransformation activity, which hinders the activation of Ha-Ras V12 -Wnt5a-Stat3 pathway.
Altogether, these results suggest that both decreasing Cav1 and increasing exosomal Wnt5a must be implemented during Ha-Ras V12 -driven cell transformation. Differential sensitivity of epithelial cells to extracellular matrix in polarity establishment. Full Text Available Establishment of apical-basal polarity is crucial for epithelial sheets that form a compartment in the body, which function to maintain the environment in the compartment.
Effects of impaired polarization are easily observed in three-dimensional 3-D culture systems rather than in two-dimensional 2-D culture systems. Although the mechanisms for establishing the polarity are not completely understood, signals from the extracellular matrix ECM are considered to be essential for determining the basal side and eventually generating polarity in the epithelial cells.
These cells showed clear apical-basal polarity in 2-D cultures. In 3-D cultures, however, each cell line displayed different responses to the same ECM. In EpH4 cells , the spheroids displayed an apical-basal polarity that was opposite to that seen in the other two cell types in both ECM gels, at least during the culture period.
On the other hand, the three cell lines showed the same apical-basal polarity both in 2-D cultures and in 3-D cultures using the hanging drop method. The three lines also had similar cellular responses to ECM secreted by the cells themselves. Therefore, appropriate culture conditions should be carefully determined in advance when using various epithelial cells to analyze cell polarity or 3-D morphogenesis. Full Text Available B-1a cells are innate-like B-lymphocytes producing natural antibodies.
Activation-induced cytidine deaminase AID, a product of the Aicda gene, plays a central role in class-switch recombination and somatic hypermutation in B cells. Although a role for Aicda in B-1a cells has been suggested on the basis of experiments with knock out KO mice, whether B-1a cells express Aicda, and if so, which B-1a cell subpopulation expresses Aicda, remains unknown.
Here, we demonstrate that B-1 cells express Aicda, but at a level below that expressed by germinal center GC B cells. We previously reported that B-1a cells can be subdivided based on CD25 expression. Full Text Available Hematopoiesis is a multistage process involving the differentiation of stem and progenitor cells into distinct mature cell lineages. Here we present Haemopedia, an atlas of murine gene- expression data containing 54 hematopoietic cell types, covering all the mature lineages in hematopoiesis.
We include rare cell populations such as eosinophils, mast cells , basophils, and megakaryocytes, and a broad collection of progenitor and stem cells. We show that lineage branching and maturation during hematopoiesis can be reconstructed using the expression patterns of small sets of genes.
We also have identified genes with enriched expression in each of the mature blood cell lineages, many of which show conserved lineage-enriched expression in human hematopoiesis. We have created an online web portal called Haemosphere to make analyses of Haemopedia and other blood cell transcriptional datasets easier. This resource provides simple tools to interrogate gene- expression -based relationships between hematopoietic cell types and genes of interest.
High expression of markers of apoptosis in Langerhans cell histiocytosis. Langerhans cell histiocytosis cells showed strong expression of p53 and in some cases co- expression of Fas and Fas-L. The expression of Fas-L was significantly higher in infiltrates from patients with single-system disease. The actual The co A delicate In conclusion, these data indicate that human thymus epithelial cells express significant levels Rethinking cell -cycle-dependent gene expression in Schizosaccharomyces pombe.
Three studies of gene expression during the division cycle of Schizosaccharomyces pombe led to the proposal that a large number of genes are expressed at particular times during the S. Yet only a small fraction of genes proposed to be expressed in a cell -cycle-dependent manner are reproducible in all three published studies. In addition to reproducibility problems, questions about expression amplitudes, cell -cycle timing of expression , synchronization artifacts, and the problem with methods for synchronizing cells must be considered.
These problems and complications prompt the idea that caution should be used before accepting the conclusion that there are a large number of genes expressed in a cell -cycle-dependent manner in S. Pharmacologic suppression of target cell recognition by engineered T cells expressing chimeric T- cell receptors.
Adoptive therapy with autologous T cells expressing chimeric T- cell receptors chTCRs is of potential interest for the treatment of malignancy. To limit possible T- cell -mediated damage to normal tissues that weakly express the targeted tumor antigen Ag , we have tested a strategy for the suppression of target cell recognition by engineered T cells.
Jurkat T cells were transduced with an anti-hapten chTCR tinder the control of a tetracycline-suppressible promoter and were shown to respond to Ag-positive hapten-coated but not to Ag-negative target cells. The engineered T cells were then reacted with hapten-coated target cells at different effector to target cell ratios before and after exposure to tetracycline.
When the engineered T cells were treated with tetracycline, expression of the chTCR was greatly decreased and recognition of the hapten-coated target cells was completely suppressed. Tetracycline-mediated suppression of target cell recognition by engineered T cells may be a useful strategy to limit the toxicity of the approach to cancer gene therapy.
Full Text Available In contrast to donor T cells , natural killer NK cells are known to mediate anti-cancer effects without the risk of inducing graft-versus-host disease GvHD. In order to improve cytotoxicity against resistant cancer cells , auspicious efforts have been made with chimeric antigen receptor CAR expressing T- and NK cells. These CAR-modified cells express antigen receptors against tumor-associated surface antigens, thus redirecting the effector cells and enhancing tumor-specific immunosurveillance.
In order to control such potentially severe side effects, the insertion of suicide genes into CAR-modified effectors can provide a means for efficient depletion of these cells. In this review we summarize the data on CAR expressing NK cells focusing on the possible advantage using these short-lived effector cells and discuss the necessity of suicide switches. Furthermore, we address the compliance of such modified NK cells with regulatory requirements as a new field in cellular immunotherapy.
Full Text Available There is a revolution in the ability to analyze gene expression of single cells in a tissue. To understand this data we must comprehend how cells are distributed in a high-dimensional gene expression space. One open question is whether cell types form discrete clusters or whether gene expression forms a continuum of states. If such a continuum exists, what is its geometry? Recent theory on evolutionary trade-offs suggests that cells that need to perform multiple tasks are arranged in a polygon or polyhedron line, triangle, tetrahedron and so on, generally called polytopes in gene expression space, whose vertices are the expression profiles optimal for each task.
Here, we analyze single- cell data from human and mouse tissues profiled using a variety of single- cell technologies. We fit the data to shapes with different numbers of vertices, compute their statistical significance, and infer their tasks. We find cases in which single cells fill out a continuum of expression states within a polyhedron.
This occurs in intestinal progenitor cells , which fill out a tetrahedron in gene expression space. The four vertices of this tetrahedron are each enriched with genes for a specific task related to stemness and early differentiation. A polyhedral continuum of states is also found in spleen dendritic cells , known to perform multiple immune tasks: cells fill out a tetrahedron whose vertices correspond to key tasks related to maturation, pathogen sensing and communication with lymphocytes.
A mixture of continuum-like distributions and discrete clusters is found in other cell types, including bone marrow and differentiated intestinal crypt cells. This approach can be used to understand the geometry and biological tasks of a wide range of single- cell datasets. The present results suggest that the concept of cell type may be expanded.
In addition to discreet clusters in gene- expression space, we suggest a new possibility: a continuum of states within a. Gene expression of manganese superoxide dismutase in human glioma cells. Full Text Available Aim This study analyze the MnSOD gene expression as endogenous antioxidant in human glioma cells compared with leucocyte cells as control.
Conclusion MnSOD gene expression in human glioma cells are significantly lower than its expression in leucocytes cells. Murine cell glycolipids customization by modular expression of glycosyltransferases. Functional analysis of glycolipids has been hampered by their complex nature and combinatorial expression in cells and tissues.
We report an efficient and easy method to generate cells with specific glycolipids. In our proof of principle experiments we have demonstrated the customized expression of two relevant glycosphingolipids on murine fibroblasts, stage-specific embryonic antigen 3 SSEA-3 , a marker for stem cells , and Forssman glycolipid, a xenoantigen.
Sets of genes encoding glycosyltansferases were transduced by viral infection followed by multi-color cell sorting based on coupled expression of fluorescent proteins. Chemokine receptor expression by inflammatory T cells in EAE. Chemokines direct cellular infiltration to tissues, and their receptors and signaling pathways represent targets for therapy in diseases such as multiple sclerosis MS. The CCL We have assessed Our approach was to induce EAE, and then examine chemokine receptor expression by cytokine-producing T cells sorted from CNS at peak disease.
During development intense Sox2 expression marks not only Prox1- expressing taste bud cell but also perigemmal cell lineages. Sox2 is proposed to regulate the differentiation of bipotential progenitor cells into taste bud cells. However, detailed expression of Sox2 remains unclear. In this report, Sox2 expression during taste bud development in the fungiform FF , circumvallate CV and soft palate SP areas is examined together with Prox1. First, we immunohistochemically checked Prox1 expression in adults and found that almost all taste bud cells are Prox1-positive.
During FF development, intense Sox2 expression was restricted to taste bud primordia expressing Prox1 at E However, at E In the SP, at E However, intense Sox2 expression was not restricted to taste bud primordia but was detected widely in the epithelium. During development, Sox2 expression outside developing taste buds was generally down-regulated but was retained in the perigemmal region similarly to that in the FF.
In the CV, the initial stage of taste bud development remained unclear because of the lack of taste bud primordia comparable to that in the FF and SP. Here, we show that Prox1- expressing cells appear in the apical epithelium at E Sox2 was again not restricted to developing taste bud cells expressing Prox1 during CV development.
The expression patterns support that Sox2 does not serve as a cell fate selector between taste bud cells and surrounding keratinocytes but rather may contribute to them both. Expression and significance of Axin2 in pancreatic cancer cells. Full Text Available ObjectiveTo investigate the expression of Axin2 in pancreatic cancer cells , and to observe the influence of Axin2 on the proliferation, invasion, and migration of human pancreatic cancer cells PANC PANC-1 cells with low expression were transfected with over- expressed Axin2 plasmid by transient transfection.
MTT assay, Transwell assay, and scratch assay were used to determine the proliferation, invasion, and migration of cells transfected with over- expressed Axin2. One-way analysis of variance was used for comparison between multiple groups, and SNK-q test was used for comparison between any two groups. Compared with those in the non-transfection group and the blank group, PANC-1 cells in the over- expression group showed significant reductions in the proliferation, invasion, and migration abilities.
ConclusionThe expression of Axin2 is down-regulated in pancreatic cancer cell lines and decreases with the increasing invasion ability, suggesting the role of tumor suppressor gene. High expression of Axin2 can reduce the proliferation, invasion, and migration abilities of PANC-1 cells. Differential marker expression by cultures rich in mesenchymal stem cells. Background Mesenchymal stem cells have properties that make them amenable to therapeutic use.
However, the acceptance of mesenchymal stem cells in clinical practice requires standardized techniques for their specific isolation. To date, there are no conclusive marker s for the exclusive isolation of mesenchymal stem cells. Our aim was to identify markers differentially expressed between mesenchymal stem cell and non-stem cell mesenchymal cell cultures.
We compared and contrasted the phenotype of tissue cultures in which mesenchymal stem cells are rich and rare. By initially assessing mesenchymal stem cell differentiation, we established that bone marrow and breast adipose cultures are rich in mesenchymal stem cells while, in our hands, foreskin fibroblast and olfactory tissue cultures contain rare mesenchymal stem cells.
In particular, olfactory tissue cells represent non-stem cell mesenchymal cells. Subsequently, the phenotype of the tissue cultures were thoroughly assessed using immuno-fluorescence, flow-cytometry, proteomics, antibody arrays and qPCR. Results Our analysis revealed that all tissue cultures, regardless of differentiation potential, demonstrated remarkably similar phenotypes.
Importantly, it was also observed that common mesenchymal stem cell markers, and fibroblast-associated markers, do not discriminate between mesenchymal stem cell and non-stem cell mesenchymal cell cultures.
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Panneerselvam K, Freeze HH. Mannose enters mammalian cells using a specific transporter that is insensitive to glucose. Stuart RO, et al. Dependence of epithelial intercellular junction biogenesis on thapsigargin-sensitive intracellular calcium stores.
Grindstaff KK, et al. Nucleotide GenBank : D Canis familiaris interleukin-8 gene, partial sequence. More Less There are no consistent identifiable marker chromosomes. Add To Cart. Permits Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Madin and N. Subculturing Volumes are given for a 75 cm 2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Remove and discard culture medium. Rinse the cell layer twice with 0.
Add 2. Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Add 6. Add appropriate aliquots of the cell suspension to new culture vessels. PubMed: Haass C, et al. PubMed: Gaush CR, et al. PubMed: Loffler S, et al. PubMed: Mead JR, et al. PubMed: von Dippe P, et al. PubMed: Stuart RO, et al.
PubMed: Grindstaff KK, et al. Documentation Permits These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. Product Sheet. Certificate of Analysis. Cell Micrograph. Login Please enter a username. Please enter a password. Invalid username or password. Canis familiaris , dog. Culture Properties. This cell line is a suitable transfection host and is useful for influenza research.
For the season, the viruses provided to the manufacturer to be grown in cell culture are cell-derived rather than egg-derived. Growing flu viruses in eggs can introduce changes called egg-adapted changes that can cause differences between the viruses in the vaccine and the ones that are circulating.
Observational studies have shown greater protection against flu or flu-like illness among people who received Flucelvax compared to those who received standard-dose egg-based vaccines. A potential advantage of cell culture technology is that it might permit faster start-up of the vaccine manufacturing process in the event of a pandemic.
Growing the flu viruses in cell culture for the manufacture of Flucelvax Quadrivalent is not dependent on an egg supply. Cell-based flu vaccines that are produced using CVVs have the potential to be more effective than traditional egg-based flu vaccines. A clinical trial of the previous trivalent formulation of Flucelvax demonstrated effectiveness and safety among persons 18 through 49 years old.
In immunogenicity studies among people 18 years and older and 4 through 17 years old, Flucelvax Quadrivalent was found to produce a similar immune response to the trivalent formulation. Post-vaccination symptoms were typical of those seen with other injectable flu vaccines. Cell culture technology has been used to produce other U. Skip directly to site content Skip directly to page options Skip directly to A-Z link.
Influenza Flu. Section Navigation. Facebook Twitter LinkedIn Syndicate. Minus Related Pages. Get a Flu Vaccine. On This Page. What is the significance of FDA approving cell-based candidate vaccine viruses for use in the Flucelvax Quadrivalent cell-based flu vaccines? What are the possible benefits of using cell-based flu vaccines?
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