The Experts below are selected from a list of 1365 Experts worldwide ranked by ideXlab platform
Julie T Daniels - One of the best experts on this subject based on the ideXlab platform.
-
characterisation and functional features of a spontaneously Immortalised human corneal epithelial Cell Line with progenitor like characteristics
Brain Research Bulletin, 2010Co-Authors: Maria Notara, Julie T DanielsAbstract:In this study a spontaneously formed corneal epithelial Cell Line, namely HCE-S, was established and characterised. The Cell Line was karyotyped and corneal epithelial maker expression of the Cell Line was assessed by immunostaining and semi-quantitative RT-PCR. The morphological characteristics were investigated using SEM and TEM analyses. The functional response to EGF in terms of Cell proliferation, wound healing and Cell migration was tested using Alamar Blue, scratch wound and colony dispersion assays, respectively. The Cells were maintained in culture for more than 100 divisions and 35 passages suggesting that an Immortalised Cell Line had been established. HCE-S, has maintained an epithelial morphology and has not phenotypicaly changed through passages. SEM and TEM microscopy showed morphological similarities to primary corneal epithelial Cells. HCE-S expressed a battery of characteristic markers of primary corneal epithelial Cells including cytokeratin 3 and PAX 6 as well as the basal Cell integrins β1 and α9 and the putative corneal stem Cell marker ABCG2. HCE-S Cells were responsive to exogenous EGF as shown by proliferation, migration and scratch wound assays. HCE-S can be cultured in a simple DMEM and only serum-based media which gives them an advantage against available corneal epithelial Cell Lines. This fact, along with the often limited availability and variability of primary corneal epithelial Cells and the similarities of the Cell Line with primary Cell characteristics suggest that HCE-S could be a useful tool for the study of corneal epithelial Cell biology, ocular surface toxicity studies and pharmacological testing.
Maria Notara - One of the best experts on this subject based on the ideXlab platform.
-
characterisation and functional features of a spontaneously Immortalised human corneal epithelial Cell Line with progenitor like characteristics
Brain Research Bulletin, 2010Co-Authors: Maria Notara, Julie T DanielsAbstract:In this study a spontaneously formed corneal epithelial Cell Line, namely HCE-S, was established and characterised. The Cell Line was karyotyped and corneal epithelial maker expression of the Cell Line was assessed by immunostaining and semi-quantitative RT-PCR. The morphological characteristics were investigated using SEM and TEM analyses. The functional response to EGF in terms of Cell proliferation, wound healing and Cell migration was tested using Alamar Blue, scratch wound and colony dispersion assays, respectively. The Cells were maintained in culture for more than 100 divisions and 35 passages suggesting that an Immortalised Cell Line had been established. HCE-S, has maintained an epithelial morphology and has not phenotypicaly changed through passages. SEM and TEM microscopy showed morphological similarities to primary corneal epithelial Cells. HCE-S expressed a battery of characteristic markers of primary corneal epithelial Cells including cytokeratin 3 and PAX 6 as well as the basal Cell integrins β1 and α9 and the putative corneal stem Cell marker ABCG2. HCE-S Cells were responsive to exogenous EGF as shown by proliferation, migration and scratch wound assays. HCE-S can be cultured in a simple DMEM and only serum-based media which gives them an advantage against available corneal epithelial Cell Lines. This fact, along with the often limited availability and variability of primary corneal epithelial Cells and the similarities of the Cell Line with primary Cell characteristics suggest that HCE-S could be a useful tool for the study of corneal epithelial Cell biology, ocular surface toxicity studies and pharmacological testing.
N. Joan Abbott - One of the best experts on this subject based on the ideXlab platform.
-
Functional expression of P-glycoprotein in an Immortalised Cell Line of rat brain endothelial Cells, RBE4
Journal of Neurochemistry, 1996Co-Authors: David J. Begley, Delphine Lechardeur, Zheng Duan Chen, Christopher Rollinson, Michèle Bardoul, Françoise Roux, Daniel Scherman, N. Joan AbbottAbstract:The presence of P-glycoprotein in the Cell plasma membrane limits the penetration of many cytotoxic substances into Cells that express the gene product. There is considerable evidence also to indicate that P-glycoprotein is expressed as part of the normal blood-brain barrier in the luminal membranes of the cerebral capillary endothelial Cells, where it presumably performs a protective function for the brain. This report describes the functional expression of P-glycoprotein in an Immortalised Cell Line, RBE4, derived from rat cerebral capillary endothelial Cells. The expression of P-glycoprotein is demonstrated by western immunoblotting and by immunogold and fluorescent staining with monoclonal antibodies. The Cellular accumulation of [3H]colchicine and [3H]-vinblastine is investigated and shown to be enhanced by the presence of azidothymidine, chlorpromazine, verapamil, cyclosporin A, and PSC 833 ([3'-keto-Bmt1]-[Val2]-cyclosporin) at 50 or 100 microM concentration. It is concluded that the RBE4 Cell Line is a valuable tool for investigating the mechanisms of P-glycoprotein activity both in the blood-brain barrier and in multidrug resistance in general.
David J. Begley - One of the best experts on this subject based on the ideXlab platform.
-
Functional expression of P-glycoprotein in an Immortalised Cell Line of rat brain endothelial Cells, RBE4
Journal of Neurochemistry, 1996Co-Authors: David J. Begley, Delphine Lechardeur, Zheng Duan Chen, Christopher Rollinson, Michèle Bardoul, Françoise Roux, Daniel Scherman, N. Joan AbbottAbstract:The presence of P-glycoprotein in the Cell plasma membrane limits the penetration of many cytotoxic substances into Cells that express the gene product. There is considerable evidence also to indicate that P-glycoprotein is expressed as part of the normal blood-brain barrier in the luminal membranes of the cerebral capillary endothelial Cells, where it presumably performs a protective function for the brain. This report describes the functional expression of P-glycoprotein in an Immortalised Cell Line, RBE4, derived from rat cerebral capillary endothelial Cells. The expression of P-glycoprotein is demonstrated by western immunoblotting and by immunogold and fluorescent staining with monoclonal antibodies. The Cellular accumulation of [3H]colchicine and [3H]-vinblastine is investigated and shown to be enhanced by the presence of azidothymidine, chlorpromazine, verapamil, cyclosporin A, and PSC 833 ([3'-keto-Bmt1]-[Val2]-cyclosporin) at 50 or 100 microM concentration. It is concluded that the RBE4 Cell Line is a valuable tool for investigating the mechanisms of P-glycoprotein activity both in the blood-brain barrier and in multidrug resistance in general.
Patricio Soaresdasilva - One of the best experts on this subject based on the ideXlab platform.
-
l dopa transport properties in an Immortalised Cell Line of rat capillary cerebral endothelial Cells rbe 4
Brain Research, 1999Co-Authors: Pedro Gomes, Patricio SoaresdasilvaAbstract:Abstract The present study aimed to determine the kinetics of l -3,4-dihydroxyphenylalanine ( l -DOPA) uptake in an Immortalised Cell Line of rat capillary cerebral endothelial Cells (clones RBE 4 and RBE 4B), to define the type of inhibition produced by l -5-hydroxytryptophan ( l -5-HTP), 2-aminobicyclo(2,2,1)-heptane-2-carboxylic acid (BHC) and N -(methylamino)-isobutyric acid (MeAlB) and its sodium dependence. Non-Linear analysis of the saturation curves for l -DOPA and l -5-HTP revealed in RBE 4 Cells K m values (in μM) of 72 and 102 and in RBE 4B Cells K m values (in μM) of 60 and 118, respectively. IC 50 values for l -5-HTP (RBE 4, 1026 μM; RBE 4B, 831 μM) obtained in the presence of a nearly saturating (250 μM) concentration of l -DOPA were almost 5-fold those obtained when non-saturating (25 μM) concentrations of l -DOPA were used. IC 50 values for BHC obtained in the presence of a nearly saturating (250 μM) concentration of l -DOPA were also 6- to 5-fold those obtained when non-saturating (25 μM) concentrations of l -DOPA were used. MeAlB (up to 2.5 mM) was found not to interfere with the uptake of l -DOPA. In RBE 4 Cells, V max values for l -DOPA uptake were identical in the absence and the presence of 150 μM l -5-HTP or 150 μM BHC, but K m values (μM) were significantly greater ( P l -DOPA uptake was studied in the presence of l -5-HTP or BHC. Similar findings were observed when RBE 4B Cells were used. Uptake of (250 μM) l -DOPA in the absence of sodium in the incubation medium was similar to that observed in the presence of increasing concentrations of sodium (20 to 140 mM). It is concluded that RBE 4 and RBE 4B Cells are endowed with the L-type amino acid transporter through which l -DOPA and l -5-HTP can be taken up, and suggested that this Immortalised Cell Line of rat capillary cerebral endothelium might constitute an interesting in vitro model for the study of BBB mechanisms, namely those concerning solute and nutrient transfer across the brain capillary endothelium.
