Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer

Bledi, Yaniv; Domb, Abraham J.; Linial, Michal

doi:10.1016/s1385-299x(00)00024-6

Cited by 50 publications

(36 citation statements)

References 22 publications

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“…To improve adhesion of this cell line to plates, the wells were treated for 40 min at room temperature with a novel polyethyleneimine-based polymer (AB-30, 12.5 g/ml in ethanol). The polymer was synthesized by Dr. W. J. Dunn III (Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, IL) according to the procedure of Bledi et al (2000). For the trafficking experiment, cells were incubated with dopamine at 37°C in serum-free medium for 1 h. The culture medium was removed from each well in the plate.…”

Section: Dat-expressing Cellsmentioning

confidence: 99%

Substrate-Induced Trafficking of the Dopamine Transporter in Heterologously Expressing Cells and in Rat Striatal Synaptosomal Preparations

Chi

Reith²

2003

J Pharmacol Exp Ther

108

100

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Dopamine transporter (DAT) trafficking was assessed by functional measurements of dopamine uptake and by biotinylation of surface proteins followed by gel electrophoresis and Western blotting. In human embryonic kidney (HEK)-293 cells expressing human DAT (HEK-hDAT), pretreatment with dopamine (0.1-100 M) followed by washout caused reductions in subsequent dopamine uptake (reflected in V max ) with effective dopamine concentrations in the 10 to 100 M range and pretreatment times of 10 to 60 min. Reductions assessed after 60-min pretreatment with 100 M dopamine corresponded with decreases measured in surface DAT by the noncleavable biotin method, which were caused, at least in part, by enhanced endocytosis as monitored with cleavable biotin. Pretreatment of rat striatal synaptosomes with dopamine (10 and 100 M) also caused reductions in DAT uptake activity (V max ), and again the underlying mechanism seemed to be a diminished presence of DAT at the surface of synaptosomes as measured by the noncleavable biotin method. The copresence of cocaine during pretreatment with dopamine prevented the down-regulation of surface DAT. The present results show that DAT surface residency can be regulated by substrate acting on it, not only in cells heterologously expressing DAT but also in situ in rat brain tissue.In the brain, the dopamine transporter (DAT) clears extraneuronal dopamine, thereby terminating dopamine action (Iversen, 1971). It is generally thought that the local density of DAT is one factor that determines how much dopamine can be cleared, just as the density of dopamine receptors is crucial for the intensity of dopamine receptor-mediated signaling. In this context, many studies over the past decade have addressed long-term regulation of DAT density in chronic drug studies (for reviews, see Kuhar and Pilotte, 1996;Zahniser and Doolen, 2001). The idea that DAT is also susceptible to short-term regulation on a scale of minutes, through changes in its presence at the cell surface, is more recent and originates in studies searching for a link between transporter activity and phosphorylation states. Thus, depending on the mode of treatment, the protein kinase C (PKC) activator ␤-phorbol 12-myristate 13-acetate (PMA) rapidly increases (Quick et al., 1997) or decreases (Beckman et al., 1999) surface-resident GABA transporter 1 (GAT1) in oocytes or primary hippocampal cultures. Down-regulation of GAT1 is probably the physiologically relevant effect of PKC activation (Robinson, 2002). In addition, phorbol esters down-regulate, or reduce surface activity, of monoamine transporters for serotonin (Qian et al., 1997), norepinephrine (Apparsundaram et al

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Section: Dat-expressing Cellsmentioning

confidence: 99%

Substrate-Induced Trafficking of the Dopamine Transporter in Heterologously Expressing Cells and in Rat Striatal Synaptosomal Preparations

Chi

Reith²

2003

J Pharmacol Exp Ther

108

100

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“…[24] In recent investigations, other polymers were surface modified with PETIM as well, which improved attachment and proliferation of neuronal cells. [25,26] Therefore, we were interested whether a PEI membrane modified with PETIM is suitable as a culture substratum for keratinocytes. A polyacrylonitrile (PAN) membrane was used for comparison because this material also promotes adhesion and growth of a variety of cells, [12,22,27] but is much less stable than PEI, which limits its clinical application.…”

Section: Introductionmentioning

confidence: 99%

Poly(ether imide) Membranes Modified with Poly(ethylene imine) as Potential Carriers for Epidermal Substitutes

Trimpert

Boese

Albrecht

et al. 2006

Macromolecular Bioscience

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Poly(ether imide) (PEI) membranes were modified with a linear low-molecular weight (PETIM_0.6) and a branched high-molecular weight poly(ethylene imine) (PETIM_60). The membrane surfaces became more hydrophilic and the zeta potentials were shifted from negative to positive zeta values after immobilisation of both PETIM. These measurements also indicated the presence of a swollen surface layer in the case of PETIM_60, while a regular structuring of the surface was observed with scanning force microscopy for PETIM_0.6. A human keratinocyte cell line HaCaT was cultured on the different membranes. It was found that HaCaT cell growth was stimulated by PETIM_0.6. Cells reached earlier confluence on this substratum, while their growth was inhibited on a PEI membrane modified with PETIM_60, which makes PEI membranes modified with PETIM_0.6 a promising material for in vitro culture of epidermal transplants.

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“…A number of explanations however, can be offered to elucidate such interactions. Cellular adhesion has been investigated on a wide variety of substrates coated with natural (poly L-lysine, fibronectin, vitronectin, collagen) and synthetic polymers (polyethylenimine) to promote cell binding [32][33][34][35]. In these cases, cellular adhesion is thought to be mediated by a variety of interactions, the most common being electrostatic interaction; wherein cellular immobilisation is promoted by a highly charged surface [25].…”

Section: Discussionmentioning

confidence: 99%

A microarray approach to the identification of polyurethanes for the isolation of human skeletal progenitor cells and augmentation of skeletal cell growth

et al. 2009

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Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer

Cited by 50 publications

References 22 publications

Substrate-Induced Trafficking of the Dopamine Transporter in Heterologously Expressing Cells and in Rat Striatal Synaptosomal Preparations

Substrate-Induced Trafficking of the Dopamine Transporter in Heterologously Expressing Cells and in Rat Striatal Synaptosomal Preparations

Poly(ether imide) Membranes Modified with Poly(ethylene imine) as Potential Carriers for Epidermal Substitutes

A microarray approach to the identification of polyurethanes for the isolation of human skeletal progenitor cells and augmentation of skeletal cell growth

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