High Throughput Screening Method for Identification of New Lipofection Reagents

Regelin, Anne; Fernholz, E.; Krug, Harald F.; Massing, Ulrich

doi:10.1089/10870570152488446

Cited by 2 publications

(3 citation statements)

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“…Such applications may include studies of cellular secretions in co-cultures of multiple cells, 26 studies of microenvironmental effects on stem cells, 40 single cells behaviour, 41,42 mapping global gene expression networks and drug screening. [43][44][45] Fig. 7 Fluorescent immunostaining of E-cadherin (green) and counterstaining with DAPI (blue) in conventional monolayer culture confirmed that administration of E-cadherin blocking antibody prevented the localisation of E-cadherin to sites of cell-cell interactions (arrows) (A).…”

Section: Discussionmentioning

confidence: 85%

Parylene peel-off arrays to probe the role of cell–cell interactions in tumour angiogenesis

et al. 2009

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Microenvironmental conditions impact tumour angiogenesis, but the role of cell-cell interactions in modulating the angiogenic capability of tumour cells is not well understood. We have microfabricated a peel-off cell-culture array (PeelArray) chip to spatiotemporally control interactions between tumour cells in a large array format and to analyse angiogenic factor secretion in response to these conditions. The PeelArray chip consists of a polyethylene glycol (PEG) treated glass coverslip coated with a parylene-C template that can be easily peeled off to selectively micropattern biomolecules and cells. We have designed the PeelArray chip to reproducibly deposit large uniform arrays of isolated single cells or isolated cell clusters on fibronectin features of defined surface areas. We have utilised this microfabricated culture system to study the secretion of angiogenic factors by tumour cells, in the presence or absence of cell-cell contact as controlled by micropatterning. Our results indicate that cell-cell interactions play a synergistic role in regulating the expression of angiogenic factors (i.e., vascular endothelial growth factor [VEGF] and interleukin-8 [IL-8]) in various cancer cell lines, independent of other more complex microenvironmental cues (e.g. hypoxia). Our PeelArray chip is a simple and adaptable micropatterning method that enables quantitative profiling of protein secretions and hence, a better understanding of the mechanisms by which cell-cell interactions regulate tumour cell behaviour and angiogenesis.

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Section: Discussionmentioning

confidence: 85%

Parylene peel-off arrays to probe the role of cell–cell interactions in tumour angiogenesis

et al. 2009

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“…Transfection and cytotoxicity assays can be similarly performed in multiwell plates with repeating samples to reduce measurement mistakes. Pioneers in this type of HT screening of a wide range of polymers as transfection agents have been Langer and co-workers (synthesis and transfection efficiency) and Massing and co-workers (lipofection transfection efficiency and toxicity) …”

Section: Introductionmentioning

confidence: 99%

“…Pioneers in this type of HT screening of a wide range of polymers as transfection agents have been Langer and co-workers (synthesis and transfection efficiency) 29 and Massing and co-workers (lipofection transfection efficiency and toxicity). 30 We describe here a simple and powerful combinatorial highthroughput workflow that combines polyplex formation and biological screening (Scheme 1). It starts with the automated polyplex preparation via pipetting robots and continues with a parallel and HT analysis of analytical and biological properties of size, binding affinity, stability, transfection efficiency, and toxicity.…”

Section: ■ Introductionmentioning

confidence: 99%

Parallel High-Throughput Screening of Polymer Vectors for Nonviral Gene Delivery: Evaluation of Structure–Property Relationships of Transfection

et al. 2013

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In recent years, "high-throughput" (HT) has turned into a keyword in polymer research. In this study, we present a novel HT workflow for the investigation of cationic polymers for gene delivery applications. For this purpose, various poly(ethylene imine)s (PEI) were used as representative vectors and investigated via HT-assays in a 96-well plate format, starting from polyplex preparation up to the examination of the transfection process. In detail, automated polyplex preparation, complex size determination, DNA binding affinity, polyplex stability, cytotoxicity, and transfection efficiency were performed in the well plate format. With standard techniques, investigation of the biological properties of polymers is quite time-consuming, so only a limited number of materials and conditions (such as pH, buffer composition, and concentration) can be examined. The approach described here allows many different polymers and parameters to be tested for transfection properties and cytotoxicity, giving faster insights into structure-activity relationships for biological activity.

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High Throughput Screening Method for Identification of New Lipofection Reagents

Cited by 2 publications

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Parylene peel-off arrays to probe the role of cell–cell interactions in tumour angiogenesis

Parylene peel-off arrays to probe the role of cell–cell interactions in tumour angiogenesis

Parallel High-Throughput Screening of Polymer Vectors for Nonviral Gene Delivery: Evaluation of Structure–Property Relationships of Transfection

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