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Molecular biology

standard molecular cloning; PCR, RT-PCR, Quantitative real-time PCR diagnostics; RNAi technology, production of knock-down mammalian cell lines

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Biochemistry

production of recombinant proteins; protein and protein complex purification (immunoaffinity, FPLC (Fast Pressure Liquid Chromatography), in vivo cross-linking); Western blotting; immunohistochemistry; immunofluorescence; immunoprecipitation

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Cell biology

production of transgenic mice by the pronuclear injection of cloned DNA sequences; mouse embryonic stem cell culture; gene-targeting in mouse embryonic stem cells; production of knock-out mice; standard mammalian cell culture, transfection and manipulation; production of transgenic mammalian cell lines; isolation and culture of adult mesenchymal stem cells and embryonic stem cells; molecular characterization of stem cell lines by FACS (Fluorescence Activated Cell Sorting)

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Cytology

FISH (Fluorescent In Situ Hybridisation) analysis; immunofluorescence; indirect immunostaining

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Cell toxicity tests

Colorimetric endpoint assays: MTT dye conversion test, LDH release assay
Cell cycle and apoptosis analysis by FACS
Double staining of cell nucleus for detection of live/dead cells by confocal microscopy
Kinetic analysis of cell toxicity by real-time cell electronic sensing (RTCA) based on impedance
Calculation of toxic concentrarion (TC) 0, 50 and 100 values

 

The RTCA-SP system (ACEA Biosciences, Inc., USA) utilizes an automatical and continuous electronic readout called impedance to non-invasively quantify adherent cell proliferation and viability in real-time. Cells are cultured on 96-well E-plate containing gold microelectronic sensor arrays. The interaction between cells and electrode generates impedance response that correlates linearly with cell index reflecting cell number, adherence, cell growth and viability.

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Measurement of compound permeability across biological barriers

Testing the bidirectional permeability of compounds and drug candidates across culture models of the blood-brain, intestinal and nasal barriers.

 

Schematic figure of bidirectional permeability measurement of compounds/drug candidates across culture models.

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Cell culture models of biological barriers

 Blood-brain barrier model:
     co-culture of primary brain endothelial and glial cells from rat or mouse
     triple co-culture model of primary brain endothelial cells, pericytes and glial cells from rat or mouse (syngeneic)
Intestinal model: human intestinal cells*
Intranasal model: human nasal epithelial cells*
Lung model: human lung epithelial cells*
*The three epithelial models are available as monocultures or co-cultures endothelial cells

 

Co-culture and triple culture model of blood-brain barrier.

Research and Development

  • Bioprocess design and engineering of biotherapeutics
  • Drug targeting across blood brain barrier
  • Blood-brain barrier model development

Consultancy

  • Biotechnology and bioengeneering
  • Blood-brain barrier permeability and drug targeting
  • Toxicity testing on mammalian and human cells using 5 different methods

Activities

    Projects

    • National Technology Programme - 1. Life sciences subprogram– DKMACTER 2009-2012.: Tumour therapy with a combined artificial chromosome-stem cell treatment
    • TAMOP-4.2.1/B-09/1/KONV-2010-0005: In vitro cell toxicity and permeability measurements
    • TAMOP-4.2.1/B-09/1/KONV-2010-0005: Effects of acute and chronic stress on the blood-brain-barrier