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Study of the effect of zinc oxide nanoparticles on cell migration
Kutějová, Monika ; Zumberg, Inna (referee) ; Chmelíková, Larisa (advisor)
Zinc oxide nanoparticles (ZnO NPs) are widely used across various industries due to their properties, and with the development of nanomaterials, they are also finding applications in medicine. This bachelor’s thesis investigates the cytotoxic effects of these nanoparticles on cultured cell lines. The study focused on the human osteosarcoma cell line Saos-2 and the fibroblast cell line NIH-3T3. During in vitro experiments, the cells were exposed to different concentrations of ZnO NPs. The cytotoxicity of each concentration was assessed using the MTT assay, which provided information on cell viability. Based on the MTT assay results, appropriate concentrations of ZnO NPs were selected for conducting a Scratch assay, with the highest concentration corresponding to the determined inhibitory concentration for NIH-3T3. The cytotoxic effects were thus evaluated by studying cell migration. Subsequent statistical analysis determined the extent of cytotoxic effects of ZnO NPs depending on the concentration and cell type.
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Monitoring of mesenchymal stem cell migration in the extracellular matrix
Zumberg, Inna ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
This master’s thesis contains a description of mesenchymal stem cells (MSC). The work includes knowledge about the process of migration and differentiation of MSCs. The theoretical part of this thesis also deals with the problem of cell cultivation in 2D and 3D environments. The most used materials for creating 3D scaffolds are described here. The practical part contains the description of the cell culture protocol for passaging and also describes the experiment in the cell laboratory. The results of the experiment are discussed using confocal microscope images. The proposed experiment was tested with a sufficient number of repetitions. The processing of microscopic images was performed in the MATLAB programming environment.
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Application of fluorescence staining for the "Scratch Wound Healing Assay" experiment
Zumberg, Inna ; Svoboda, Ondřej (referee) ; Chmelíková, Larisa (advisor)
This bachelor’s thesis studies the cell culture of 3T3 cell line (mouse embryonic fibroblast cells) and cell migration. The Scratch Wound Healing Assay was used for the study of cell migration. The basic principles of experimenting, its advantages and disadvantages, as well as the practical use of this method in scientific research were presented on specific examples. Practical part of the bachelor’s thesis describes the experiment in the cell laboratory including cell culture of 3T3 cell line. The proposed experiment was tested with a sufficient number of repetitions. The statistical processing of microscopic images was performed in the MATLAB programming environment.
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Evaluation of antibiotic-induced mitochondrial superoxide production in adherent cells
Ingrová, Kateřina ; Chmelíková, Larisa (referee) ; Zumberg, Inna (advisor)
The theoretical part of this bachelor´s thesis contains a description of the effect of reactive oxygen species on oxidative stress by mitochondria and the consequences of antibiotics use in cell line culturing. The cell line studied in this bachelor´s thesis is the mesenchymal stem cells (MSCs). The practical part describes the procedure of the experiment including cell culturing, passaging and cell labeling. The proposed experiment was repeated with sufficient number of repetitions. Finally, confocal microscopy images were processed in the MATLAB programming environment.
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Water quality analysis using colorimetry
Kovaldová, Eliška ; Zumberg, Inna (referee) ; Čmiel, Vratislav (advisor)
This bachelor's thesis deals with the analysis of water quality using colorimetry. Part of the work is an introduction to the physical and chemical parameters of water, as well as optical methods, which include photometry, spectrophotometry and colorimetry, and an introduction to regression analysis. These methods and analysis are followed by a practical part by measuring pH, nitrate and water hardness using a spectrophotometer and an AS7262 visible light sensor with a SparkFun development board. At the end of the work there is a comparison of absorption spectra, calibration curves and linear regression parameters of both methods.
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Mitochondria morphology and membrane potential under stress conditions
Ezati, Masoumeh ; Zumberg, Inna ; Cmiel, Vratislav
Mitochondria, organelles found in the cytoplasm of almost all eukaryotic cells, generate large quantities of energy in the form of adenosine triphosphate (ATP). In addition to producing energy, mitochondria store calcium for cell signaling activities and play an important role in maintenance of ionic balance. Mitochondria, however, are highly sensitive to any kind of stress in which they mainly response by disturbance of respiration, reactive oxygen species (ROS) production and release of cytochrome c into the cytoplasm. Osmotic stress, in particular, generates ROS that degrade lipids, proteins, and DNA. High levels of salt concentration can cause an imbalance in cellular ion homeostasis that results in ion toxicity and osmotic stress. This study aims to investigate possible effects of KCl and NaCl on Bone marrow-derived (MSCs) mitochondria membrane potential (MMP) and morphology. The results indicated that KCl and NaCl of salt concentration can cause an imbalance in cellular ion homeostasis that results in ion toxicity and osmotic stress. This study aims to investigate possible effects of KCl and NaCl on MSCs mitochondria membrane potential (MMP) and morphology. The results indicated that KCl and NaCl decreased the potential and changed the morphology of mitochondria membrane compared to cells growing in normal condition.
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Evaluation of antibiotic-induced mitochondrial superoxide production in adherent cells
Ingrová, Kateřina ; Chmelíková, Larisa (referee) ; Zumberg, Inna (advisor)
The theoretical part of this bachelor´s thesis contains a description of the effect of reactive oxygen species on oxidative stress by mitochondria and the consequences of antibiotics use in cell line culturing. The cell line studied in this bachelor´s thesis is the mesenchymal stem cells (MSCs). The practical part describes the procedure of the experiment including cell culturing, passaging and cell labeling. The proposed experiment was repeated with sufficient number of repetitions. Finally, confocal microscopy images were processed in the MATLAB programming environment.
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Monitoring The Parameters Of Cells Migrating In Pseudo-3d Extracellular Matrix
Zumberg, Inna
Cell migration plays an essential role in a number of biological processes, such as embryogenesis,the immune response, wound healing and inflammation. Most research regarding cellmigration is based on experiments using two-dimensional (2D) cell cultures, and the detailed molecularand biophysical mechanisms of these processes are already well known. However, muchless is known about cell behaviour in the three-dimensional (3D) environment of living tissues.Pseudo-three-dimensional (pseudo-3D) cell cultures bridge the gap between 2D and 3D geometriesand combine their advantages. This review presents the quantitative evaluation of human mesenchymalstem cells (hMSCs) migration parameters in pseudo-3D artificial extracellular matrix(ECM) of various compositions.
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Monitoring of mesenchymal stem cell migration in the extracellular matrix
Zumberg, Inna ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
This master’s thesis contains a description of mesenchymal stem cells (MSC). The work includes knowledge about the process of migration and differentiation of MSCs. The theoretical part of this thesis also deals with the problem of cell cultivation in 2D and 3D environments. The most used materials for creating 3D scaffolds are described here. The practical part contains the description of the cell culture protocol for passaging and also describes the experiment in the cell laboratory. The results of the experiment are discussed using confocal microscope images. The proposed experiment was tested with a sufficient number of repetitions. The processing of microscopic images was performed in the MATLAB programming environment.
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Application of fluorescence staining for the "Scratch Wound Healing Assay" experiment
Zumberg, Inna ; Svoboda, Ondřej (referee) ; Chmelíková, Larisa (advisor)
This bachelor’s thesis studies the cell culture of 3T3 cell line (mouse embryonic fibroblast cells) and cell migration. The Scratch Wound Healing Assay was used for the study of cell migration. The basic principles of experimenting, its advantages and disadvantages, as well as the practical use of this method in scientific research were presented on specific examples. Practical part of the bachelor’s thesis describes the experiment in the cell laboratory including cell culture of 3T3 cell line. The proposed experiment was tested with a sufficient number of repetitions. The statistical processing of microscopic images was performed in the MATLAB programming environment.
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