|
|
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.
|
|
|
Mobile Switching Centre Emulator
Králíček, Jan ; Ráb, Jaroslav (referee) ; Ščuglík, František (advisor)
This thesis attempts to provide basic overview of the topic of the GSM system and tries to design Mobile Switching Centre Emulator that could perform the operation Location Update. The first part of this thesis describes the principles and specifics of the GSM network (network structure, the operations required to support user mobility, network entities, etc.). The second part of this thesis describes the SS7 signaling protocols, SIGTRAN and MAP protocol. The final part deals with design of the emulator.
|
|
|
Study of mesenchymal stem cell migration based on principles of chemotaxis
Pošustová, Veronika ; Skopalík, Josef (referee) ; Chmelíková, Larisa (advisor)
The purpose of this Master thesis is to verify migration of mesenchymal stem cells on the principle known as chemotaxis. First part of this study is focused on cell migration in order to explain the whole migration process. Next part describes various chemotaxis methods and selected studies dealing with clinical applications of mesenchymal stem cells in different medical and biomedical fields. The following step describes confocal microscopy, which is used for acquiring images of the cells. The experimental part is focused on cultivation of mesenchymal stem cells in a laboratory, which is necessary for cell vitality. Furthermore, there are designed two main experiments. Firstly there is a 2D experiment with adherent cells for chemotaxis using -Slide Chemotaxis. Secondly Transwell migration test is designed and executed. Finally, the acquired images from confocal microscope are used for image processing, which was done in Matlab R2020a programming environment. The result of this processing is evaluation of cell confluence and migration. In the end, experimental part of this study was optimized according to recommended studies. The results are summarized in the conclusion with proposal for improvements of those methods.
|
|
|
Construction and evaluation of in vitro set of holes for chondrogenic differentiation of MSC cells
Kříž, Marek ; Sekora, Jiří (referee) ; Čmiel, Vratislav (advisor)
Bachelor thesis concentrates on the cultivation of mesenchymal cells in vitro, damage to the articular cartilage and its replacement with the chondrogenic cells. The next section is devoted to each type of temperature control for heating and maintaining the temperature inside a closed system, the type of detector temperature and heating using the Peltier element. Assembly heater box, measuring the temperature profile inside the box with Labview program, control with Arduino platform, survival and differentiation of cells.
|
|
|
Analysis of Parkinson's disease using segmental speech parameters
Mračko, Peter ; Mekyska, Jiří (referee) ; Smékal, Zdeněk (advisor)
This project describes design of the system for diagnosis Parkinson’s disease based on speech. Parkinson’s disease is a neurodegenerative disorder of the central nervous system. One of the symptoms of this disease is disability of motor aspects of speech, called hypokinetic dysarthria. Design of the system in this work is based on the best known segmental features such as coefficients LPC, PLP, MFCC, LPCC but also less known such as CMS, ACW and MSC. From speech records of patients affected by Parkinson’s disease and also healthy controls are calculated these coefficients, further is performed a selection process and subsequent classification. The best result, which was obtained in this project reached classification accuracy 77,19%, sensitivity 74,69% and specificity 78,95%.
|
|
|
Analyser of DMX512 Protocol with USB Interface
Bartoň, Zdeněk ; Hošek, Jiří (referee) ; Kouřil, Jiří (advisor)
Aim of this work is to make know readers with modern technologies for driving scenic and musical techniques, with focus especially on a new technologies of communication. Subsequently this work deals with proposal and realization electronic device enabling analysis of control protocol DMX512, but also testing bus DMX512.
|
|
|
Construction and evaluation of in vitro set of holes for chondrogenic differentiation of MSC cells
Kříž, Marek ; Sekora, Jiří (referee) ; Čmiel, Vratislav (advisor)
Bachelor thesis concentrates on the cultivation of mesenchymal cells in vitro, damage to the articular cartilage and its replacement with the chondrogenic cells. The next section is devoted to each type of temperature control for heating and maintaining the temperature inside a closed system, the type of detector temperature and heating using the Peltier element. Assembly heater box, measuring the temperature profile inside the box with Labview program, control with Arduino platform, survival and differentiation of cells.
|
|
|
Preparation and characterisation of mRNA/DNA transfection vectors
Horák, Tomáš ; Chmelíková, Larisa (referee) ; Skopalík, Josef (advisor)
This diploma thesis deals with genetic engineering, especially the transfection of DNA into MSCs (Mesenchymal stromal cells) and dendritic cells. Both lipoplexes and metal magnetic nanoparticles were tested to introduce the vector into cells. The research was focused on finding more efficient methods of transfection. According to analysis on MADLS and gel electrophoresis, aspects playing an important role in conjugation and subsequent transfection were found. Conjugation occurs after only 4 minutes, as evidenced by an increase in zeta potential, but to achieve full conjugation it is necessary to incubate the sample for 20 minutes. Incomplete conjugation to iron nanoparticles resulted in strong carrier-carrier interactions, which formed an unwanted conglomerates. Encapsulation into liposomes with cationic surface treatment was without complications. The success rate of GFP-labeled protein expression after transfection by these methods was calculated to be 95%, resp. 91%. This result is due to low cytotoxicity. However, commercial tested kits on dendritic cells had a success rate below 5% with high cytotoxicity.
|
|
|
Preparation and characterisation of mRNA/DNA transfection vectors
Horák, Tomáš ; Chmelíková, Larisa (referee) ; Skopalík, Josef (advisor)
This diploma thesis deals with genetic engineering, especially the transfection of DNA into MSCs (Mesenchymal stromal cells) and dendritic cells. Both lipoplexes and metal magnetic nanoparticles were tested to introduce the vector into cells. The research was focused on finding more efficient methods of transfection. According to analysis on MADLS and gel electrophoresis, aspects playing an important role in conjugation and subsequent transfection were found. Conjugation occurs after only 4 minutes, as evidenced by an increase in zeta potential, but to achieve full conjugation it is necessary to incubate the sample for 20 minutes. Incomplete conjugation to iron nanoparticles resulted in strong carrier-carrier interactions, which formed an unwanted conglomerates. Encapsulation into liposomes with cationic surface treatment was without complications. The success rate of GFP-labeled protein expression after transfection by these methods was calculated to be 95%, resp. 91%. This result is due to low cytotoxicity. However, commercial tested kits on dendritic cells had a success rate below 5% with high cytotoxicity.
|
|
|
Multipotent mesenchymal stromal cells in orthopedic: Potentiation of bone healing
Stehlík, David ; Trč, Tomáš (advisor) ; Janíček, Pavel (referee) ; Sedmera, David (referee)
The aim of the thesis was development of an innovative treatment of bone defects. Human multipotent mesenchymal stromal cells (MSC) play a crucial role in bone healing. Clinical applications of MSC require large amount of cells, which could be obtained by autologous expansion of MSC harvested from bone marrow. As a first step, the standard protocol of MSC expansion based on αMEM medium and fetal bovine serum (FBS) was used. Experiments replacing FBS by pooled human serum (HS) in the culture medium concluded in patenting of a new MSC cultivation protocol (EU 1999250, CR 301141). This one-step cultivation protocol and xenogeneic protein-free cultivation medium is based on CellGro® for Hematopoietic Cells' Medium, HS, human recombinant growth factors, dexamethasone, insulin and ascorbic acid. The preclinical in vitro and in vivo experiments with MSC from both expansion protocols were carried out. Fibrillar polylactic scaffolds were seeded with MSC, cultured, differentiated and implanted in immunodeficient mice (NOD/LtSz-Rag1-). Bone-like mineralized tissue containing vessels was observed. The MSC cultured according to patented method were classified as Advanced-therapy Medicinal Product and has to fulfil the European Medicines Agency regulations to enter the clinical trials. Nevertheless the use of MSC seems...
|
guest ::
