Detection of intracellular biomarkers in viable cells using millisecond pulsed electric fields
Autor: | Justina Kavaliauskaitė, Arūnas Stirkė, Genė Biziulevičienė, Monika Petreikytė, Aida Kaušylė, Vytautas Kašėta |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Cell Membrane Permeability Cell Survival Intracellular Space Cell Count Jurkat cells Flow cytometry Cell membrane 03 medical and health sciences Jurkat Cells Mice 0302 clinical medicine Electricity medicine Animals Humans Viability assay Cells Cultured biology medicine.diagnostic_test Electroporation Cell Biology Flow Cytometry Molecular medicine Electric Stimulation Cell biology Mice Inbred C57BL 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis biology.protein Female Antibody Intracellular Biomarkers |
Zdroj: | Experimental cell research. 389(1) |
ISSN: | 1090-2422 |
Popis: | Reversible electroporation is a temporary permeabilization of cell membrane through the formation of transient pores created by short high voltage electric pulses. This method has numerous applications in biology and biotechnology and has become an important technique in molecular medicine. Reversible electroporation is usually used to transfer macromolecules into the cells. However, the delivery of large molecules such as proteins into cells without loss of cell viability remains a challenge. In our study, we investigated whether electroporation can be used for this purpose. The study was performed with the primary mouse splenocytes and Jurkat cell line. The electroporation efficacy was evaluated by flow cytometry. We used the reversible electroporation for intracellular marker detection investigating antibody and fluorescein-conjugated dextran transfer efficiency, cell viability and metabolic activity. We have found that reversible electroporation parameters can be optimized for efficient transfer of large molecules such as antibodies/proteins into live cells without a significant loss of cell viability. We conclude that a well-established and relatively easy method of reversible electroporation can be adjusted to detect intracellular biomarkers in viable cells. This is a new approach on how electroporation could be utilised in medicine and biological research to detect rare subpopulations of cells that produce specific markers and to keep cells viable. This would allow the use of these rare subpopulations of isolated cells for further research and personalized medicine. |
Databáze: | OpenAIRE |
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