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Evaluation of the effects of antibiotics on cytotoxicity of EGFP and DsRed2 fluorescent proteins used for stable cell labeling

100%

Zakrzewska K., Samluk A., Pluta K., Pijanowska D.

2014

vol. 61

issue 4

809-813

The use of fluorescent markers has proven to be an attractive tool in biological imaging. However, its usefulness may be confined by the cytotoxicity of the fluorescent proteins. In this article, for the first time, we have examined an influence of the antibiotics present in culture medium on cytotoxicity of the EGFP and DsRed2 markers used for whole-cell labeling. Results showed that doxycycline negatively affected albumin synthesis in DsRed2-expressing hepatoma cells, and that both hepatoma cells and human skin fibroblasts, labeled with this protein, were characterized by the lowered growth rates. Thus, the cytotoxic effect of fluorescent markers depends on both protein used for cell labeling and on growth conditions that may cause cell stress.

Dried human skin fibroblasts as a new substratum for functional culture of hepatic cells

76%

Wencel A., Zakrzewska K., Samluk A., Noszczyk B., Pijanowska D., Pluta K.

The primary hepatocytes culture is still one of the main challenges in toxicology studies in the drug discovery process, development of in vitro models to study liver function, and cell-based therapies. Isolated hepatocytes display a rapid decline in viability and liver-specific functions including albumin production, conversion of ammonia to urea, and activity of the drug metabolizing enzymes. A number of methods have been developed in order to maintain hepatocytes in their highly differentiated state in vitro. Optimization of culture conditions includes a variety of media formulations and supplements, growth surface coating with the components of extracellular matrix or with synthetic polymers, three-dimensional growth scaffolds and decellularized tissues, and coculture with other cell types required for the normal cell-cell interactions. Here we propose a new substratum for hepatic cells made by drying confluent human skin fibroblasts' culture. This growth surface coating, prepared using maximally simplified procedure, combines the advantages of the use of extracellular matrices and growth factors/cytokines secreted by the feeder layer cells. In comparison to the hepatoma cells grown on a regular tissue culture plastic, cells cultured on the dried fibroblasts were able to synthesize albumin in larger quantities and to form greater number of apical vacuoles. Unlike the coculture with the living feeder layer cells, the number of cells grown on the new substratum was not reduced after fourteen days of culture. This fact could make the dried fibroblasts coating an ideal candidate for the substrate for non-dividing human hepatocytes.

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2 Acta Biochimica Polonica

1 2017

1 2014

2 Pijanowska D.

2 Pluta K.

2 Samluk A.

2 Zakrzewska K.

1 Noszczyk B.

1 Wencel A.

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Evaluation of the effects of antibiotics on cytotoxicity of EGFP and DsRed2 fluorescent proteins used for stable cell labeling

Dried human skin fibroblasts as a new substratum for functional culture of hepatic cells