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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.
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Elaboration and introduction of the method for determination of some proteins
Hruzík, Ondřej ; Buňka, František (referee) ; RNDr.Karel Gebauer (advisor)
Core protein of aggrecan has a significant share on the correct function of articular cartilage. Its lack or structural failure could be the reason for the disfunction of the cartilage. The culture of chondrocytes taken from a pork articular cartilage was used for the study of aggrecan production. The monolayer culture method offers the model system which has enabled us to watch the aggrecan production into growth medium. The aggrecan synthesis was stimulated in the media with addition of L-methionin, L-serin and sodium selenite pentahydrate. Methionin and serin are antecedents of sulphur amino acid of cysteine, whose role is incredibly important for the correct function of core protein. Growth media and chondrocytes were analysed with the help of the automatic amino acids analyzer unit after acid or oxidative hydrolysis. The analyse established the amino acid representation. The main attention was paid to cysteine. The changing concentrations of this amino acid were showing if the antecedents in the addition are used for its production and, therefore, if it is possible to stimulate the production of core protein with these antecedents. The results are discussed in the conclusion of this thesis. The next step should be the detection of the concentration of synthesized aggrecan by the immunological method. Presently this method is very expensive. Therefore, the method of setting the core protein of aggrecan with the help of suitable amino acid was used for the first tests.
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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.
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Coaxial nanofibers with incorporated suplements for regulated chondrogenic differentiation
Korbelová, Gabriela ; Rampichová, Michala (advisor) ; Vištejnová, Lucie (referee)
In the field of regenerative medicine, regeneration of cartilage defects (caused either by injury or age-related degeneration) has become a widely discussed topic. Nanofibrous scaffolds provide a suitable environment for cell adhesion, proliferation, differentiation, and also for the local involvement of bioactive substances. Nanofibrous scaffolds mimic the extracellular matrix (ECM) of hyaline cartilage. These scaffolds are seeded with autologous chondrocytes. After having been isolated from the patient, the cells must be cultivated in vitro in order to obtain a sufficient amount of chondrocytes. Scaffolds with cultivated chondrocytes are later implanted back into the pacient. Chondrocytes, however, when grown on a 2D tissue culture plastic rapidly de-differentiate and thus lose the ability to synthesize ECM molecules. The aim of the work was modulation of chondrogenic differentiation medium through finding the ideal concentration of chondrogenic supplements, composed of L-ascorbate-2-phosphate (A2P) and dexamethasone (DEX), in the culture of primary chondrocytes seeded on a nanofibrous polycaprolactone (PCL) scaffold. The effect of different concentrations of the chondrogenic supplements on chondrocyte adhesion to the scaffold and their proliferation and differentiation was studied. The influence...
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Coaxial nanofibers with incorporated supplements for regulated chondrogenic differentiation
Korbelová, Gabriela ; Rampichová, Michala (advisor) ; Vištejnová, Lucie (referee)
In the field of regenerative medicine, regeneration of cartilage defects (caused either by injury or age-related degeneration, such as osteoporosis) has become a widely discussed topic. Nanofibrous scaffolds provide a suitable environment for cell adhesion, proliferation, differentiation, and also local involvement of bioactive substances. Nanofibrous scaffolds mimic the extracellular matrix (ECM) of hyaline cartilage and thus have the potential to treat cartilage defects. The aim of the work was modulation of chondrogenic differentiation medium through finding the ideal concentration of chondrogenic supplements, composed of ascorbate-2- phosphate and dexamethasone, in the culture of primary chondrocytes of pig origin seeded on a nanofibrous polycaprolactone (PCL) scaffold. The effect of different concentrations of the chondrogenic supplements on chondrocyte adhesion to the scaffold and their proliferation and differentiation was studied. Firstly, the influence of each of the supplements alone in the medium was studied, followed by study of effects of their combinations. Then, the supplements were incorporated into the nanofibers and their effect upon their release from the nanofibers was investiaged. The supplements were studied in 21-day experiments. The chondrogenic re- differentiation was best...
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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.
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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.
|
|
|
Elaboration and introduction of the method for determination of some proteins
Hruzík, Ondřej ; Buňka, František (referee) ; RNDr.Karel Gebauer (advisor)
Core protein of aggrecan has a significant share on the correct function of articular cartilage. Its lack or structural failure could be the reason for the disfunction of the cartilage. The culture of chondrocytes taken from a pork articular cartilage was used for the study of aggrecan production. The monolayer culture method offers the model system which has enabled us to watch the aggrecan production into growth medium. The aggrecan synthesis was stimulated in the media with addition of L-methionin, L-serin and sodium selenite pentahydrate. Methionin and serin are antecedents of sulphur amino acid of cysteine, whose role is incredibly important for the correct function of core protein. Growth media and chondrocytes were analysed with the help of the automatic amino acids analyzer unit after acid or oxidative hydrolysis. The analyse established the amino acid representation. The main attention was paid to cysteine. The changing concentrations of this amino acid were showing if the antecedents in the addition are used for its production and, therefore, if it is possible to stimulate the production of core protein with these antecedents. The results are discussed in the conclusion of this thesis. The next step should be the detection of the concentration of synthesized aggrecan by the immunological method. Presently this method is very expensive. Therefore, the method of setting the core protein of aggrecan with the help of suitable amino acid was used for the first tests.
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