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1

The oviduct of transgenic birds ? a source of therapeutic proteins?

100%
Bednarczyk M., Lakota P., Wawrzynska M.
Biotechnologia
|
2006
|
issue 3
134-144
EN
The use of transgenic mammals for the production of therapeutic proteins was developed over two decades ago, and a number of proteins have been produced in the milk of animals. Therapeutic proteins produced in eggs of transgenic hens have significant advantages over transgenic mammalian systems, including: regular high eggs production, large number of progeny, short generation times, sterile contents of the eggs produced by specific pathogen free chickens and, well characterization of the regulatory sequences of white protein genes similar to human glycosylation of chicken IgGs, etc. In this overview, recent progress in the development of chicken bioreaktors was presented.
2

Influence of electroporation on chicken blastoderm cell viability in vitro

100%
Wawrzynska M., Bednarczyk M., Lakota P., Lubiszewska M.
Folia Biologica
|
2008
|
vol. 56
|
issue 3-4
197-201
EN
The aim of this study was to compare two types of devices used for blastoderm cell (BC) transfection: the Nucleofector (Amaxa, Biosystems) and the Multiporator (Eppendorf). To assess the influence of electric current on BCs, different conditions of both nucleofection and electroporation were used. Next, the viability of cells was assessed. The highest number of cells (90.8%) was viable after nucleofection in the G10 program,. After transfection in the presence of pmaxGFP, the A23 program was found to be most advantageous. The elecroporation experiment with theMultiporator (Eppendorf) showed a significant influence of osmotic pressure and voltage on BC viability. Namely, in the isoosmolar buffer BC viability was statistically higher (P#0.05) in comparison to the hypoosmolar buffer. The, viability of cells was statistically higher (P#0.05) after application of 25V as compared to 50V. The efficiency of transfection in the presence of EGFP-C1 after electroporation in 2 pulses, 25V, 500 ?s in the isoosmolar buffer was better than in the recommended conditions in the Amaxa Biosystems A23 program.
3

In vitro photodynamic therapy with chlorin e6 leads to apoptosis of human vascular smooth muscle cells

76%
Wawrzynska M., Kalas W., Bialy D., Ziolo E., Arkowski J., Mazurek W., Strzadala L.
Archivum Immunologiae et Therapiae Experimentalis
|
2010
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vol. 58
|
issue 1
67-75
EN
Percutaneous coronary intervention has become the most common and widely implemented method of heart revascularization. However, the development of restenosis remains the major limitation of this method. Photodynamic therapy (PDT) recently emerged as a new and promising method for the prevention of arterial restenosis. Here the efficacy of chlorin e6 in PDT was investigated in vitro using human vascular smooth muscle cells (TG/HA-VSMCs) as one of the cell types crucial in the development of restenosis. PDT-induced cell death was studied on many levels, including annexin V staining, measurement of the generation reactive oxygen species (ROS) and caspase-3 activity, and assessment of changes in mitochondrial membrane potential and fragmentation of DNA. Photosensitization of TG/HA-VSMCs with a 170 M of chlorin e6 and subsequent illumination with the light of a 672-nm diode laser (2 J/cm2) resulted in the generation of ROS, a decrease in cell membrane polarization, caspase-3 activation, as well as DNA fragmentation. Interestingly, the latter two apoptotic events could not be observed in photosensitized and illuminated NIH3T3 fibroblasts, suggesting different outcomes of the model of PDT in various types of cells. The results obtained with human VSMCs show that chlorin e6 may be useful in the PDT of aerial restenosis, but its efficacy still needs to be established in an animal model.
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