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Stimulation of mesenchymal stem cells osteogenic differentiation using perfusion bioreactor
Šljivnjak, Erna
;
Rampichová, Michala
(advisor) ;
Rösel, Daniel
(referee)
Bone cells in vivo reside in a dynamic environment exposed to constant chemical and mechanical stimuli caused by the interstitial fluid flow. It is hypothesized that perfusion of the medium through the scaffold increases the mass transport and creates at the same time shear stress, thereby in vitro simulating the interstitial fluid effects and bone tissue formation conditions. This work examined the effects of perfusion flow rates on cell viability, proliferation, migration and osteogenic differentiation of human mesenchymal stem cells within cell-seeded 3D poly-ε-caprolactone scaffolds. Scaffolds were perfused for 21 days at flow rates 1, 3 and 5 mL/min and were compared to the scaffolds from static culture. Cells were most viable, had upregulated expression of osteogenic markers collagen type I and highest alkaline phosphatase activity under flow rate 1 mL/min when compared to their static counterparts. Cells proliferated the most under flow rate 3 mL/min when compared to their static counterparts. Flow rate 5 mL/min did not significantly differ from the static culture in any of the examined parameters. Cell migration into the scaffold was not improved upon exposure to perfusion. This data confirms that medium perfusion may benefit cell proliferation and osteogenic differentiation by enhancing...
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Evaluation of influence of mechanical loading on differentiation of stem cells into smooth muscle cells
Pražák, Šimon
;
Filová, Elena
(advisor) ;
Maxová, Hana
(referee)
Cultivation of cells in bioreactors with mechanical load simulates the physiological conditions to which cells in the body are exposed. This technology has been used to induce the differentiation of stem cells from adipose tissue towards the phenotype of vascular smooth muscle cells, which can further serve to form vascular replacements. At present, there is no established strategy for cultivating stem cells while being exposed to mechanical stress. The main aim of this work was therefore to optimize the cultivation strategy and determine the ideal load parameters. Differentiation was analyzed by immunofluorescence of specific smooth muscle cell markers, α-actin and h1-calponin, which were quantified by Western blot. Extracellular matrix production was also detected by immunofluorescence staining. The outcome of this work is the establishment of ideal conditions of cell culture in a bioreactor with mechanical load, during which they differentiate into smooth muscle cells. Three types of scaffolds were used for cultivation; plasma treated glass, fibrin-coated glass and decelularized pericardium. Preliminary results show that smooth muscle differentiation was succesfully induced in human and porcine adipose tissue stem cells. Cells were analyzed after 3 and 7 days of culture. Developing a stem cell...
Detailed record
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