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Issues of the algae Chlorella production in flow bioreactors
Jankovičová, Kristína ; Lukavský,, Jaromír (referee) ; Svěrák, Tomáš (advisor)
Microalgae invite the attention of scientists due to their unique properties, including their quick growth, accumulation of lipids and other valuable substances, fixation of carbon dioxide and treatment of wastewater. This master´s thesis is focused on the study of microalgae. The main goal is to understand and describe the process of microalgae cultivation, in order to optimize it. The theoretical part of this thesis deals with microalgae (mainly Chlorella sp.) characterization, its practical use and cultivation optimization in order to obtain the highest concentration of biomass. The experimental part is divided into three tasks. Aim of the first task was the comparison of the course of autotrophic and heterotrophic cultivation of various strains of Chlorella and Coccomyxa microalgae, using three different cultivation media – synthetic medium for chlorella cultivation and natural fertilizer, Florium, used in two different concentrations (diluted 50 and 20 times). The highest Chlorella sp. biomass concentration of 7,10 g/l was achieved in the synthetic heterotrophic medium. Second task was focused on monitoring of the growth of algae Coccomyxa and Chlorella strain C1A, with respect to temperature and light intensity, using various combinations of these two important growth factors. Chlorella achieved its highest concentration of 11,46 g/l when grown at temperature of 33,5 °C and light intensity of 320 µE.m2.s1. The third and final task was to observe the growth of Dictiosphaerium chlerelloides microalgae on a flat cascade bioreactor. The experiment led to the discovery that these algae were able to grow at temperatures of around 10 °C, at which many well-known commercial algae, such as Chlorella sp. or Arthrospina sp., simply wouldn’t grow.
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Issues of the algae Chlorella production in flow bioreactors
Jankovičová, Kristína ; Kaštánek,, Petr (referee) ; Svěrák, Tomáš (advisor)
The bachelor's thesis is focused on cultivation of Chlorella microalgae species. Theoretical part deals with microalgae characteristics, their usage and methods of their cultivation. It is focused on the most important growth factors of chlorella and compares findings from earlier studies. Experimental part contains observations of microalgae growth in different environments and behavioural comparison of autotrophic and heterotrophic cultures. Next, the influence of temperature and light intensity on microalgae growth was observed, using different combinations of these factors. Experimental findings were evaluated from gained biomass concentration and absorbance determined by spectrophotometric method.
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Mechanisms of Vascularization in Skin Tissue Engineering
Futóová, Terézia ; Brož, Antonín (advisor) ; Šuca, Hubert (referee)
Tissue engineering is a multidisciplinary field dealing with the fabrication of artificial tissue substitutes for regenerative medicine. Current regenerative medicine uses various types of tissue grafts, which have different advantages and disadvantages depending on their origin, such as insufficient amount of replacement tissue when using autologous grafts or immunogenicity of allogeneic or xenogeneic grafts. An alternative could be artificial tissue replacement. Artificial tissue constructs may consist of a non-living matrix and a cellular component. The cellular component may remodel the construct, form a functional part of the construct, or help integrate the construct into the host body. A significant problem in the formation of such replacements is sufficient vascularization. It is essential to keep cells in larger tissue constructs alive. Vascularization can be enhanced by the addition of vascular endothelial cells that can form capillaries independently within the construct. Vascular formation can also be aided by angiogenic growth factors by their direct application to the construct or by their formation, e.g. in stem cells cultured in the construct. Another approach is 3D bioprinting, allowing direct placement of specific cell types, growth factors or biomaterials in the construct. This...
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Differentiation of mesenchymal stem cells on fibrin assemblies supported by immobilized growth factors FGF2 and VEGF
Musílková, Jana ; Filová, Elena ; Kaplan, Ondřej ; Bačáková, Lucie
Bioartificial heart valves and vascular grafts prepared from decellularized tissues could be recellularized with bone marrow-derived mesenchymal stem cells (MSCs) that are able to differentiate into both smooth muscle cells and endothelial cells. MSCs differentiation is facilitated by sustained release of growth factors. In our study assemblies based on fibrin, fibrin with heparin, fibrin with adsorbed or covalently-immobilized vascular endothelial growth factor A165 (VEGF) or basic fibroblast growth factor (FGF-2) via binding to heparin attached to fibrin have been prepared and were evaluated for their stimulation of MSCs differentiation. We estimated the mRNA expression of endothelial marker CD31 (PECAM1), smooth muscle marker α-actin (ACTA2), osteoblast markers osteocalcin (BGLAP) and alkaline phosphatase (ALP). The gene expression was estimated using RT-PCR on days 1, 7 and 21 after seeding. The cell morphology and viability was evaluated by LIVE/DEAD staining. VEGF, both adsorbed and covalently bound, increased significantly the expression of smooth muscle marker α-actin. The mRNA expression of ACTA2 on day 7 and 21 raised more than 200 times in comparison to control samples (undifferentiated cells before seeding). The ACTA2 gene expression significantly exceeded the expression of all other evaluated genes at all time intervals. Moreover, on day 21, the late smooth muscle marker desmin (DES) was steeply rising in cells cultivated on assemblies containing heparin and covalently bound VEGF. The expression of osteocalcin was minimal. We conclude that fibrin assembly containing covalently bound VEGF is the most convenient for MSCs differentiation towards smooth muscle cells.
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Smart biomaterials for connective tissues regeneration
Hefka Blahnová, Veronika ; Filová, Eva (advisor) ; Maxová, Hana (referee) ; Motlík, Jan (referee)
Connective tissues are characterized by significant volume of extracellular matrix. Their main role is to provide a mechanical support and protection to other body organs. This thesis is focused on regeneration of bone, cartilage and osteochondral defect. In the experimental part we observed viability and differentiation of human mesenchymal stem cells. In vitro we evaluated the potential of PCL scaffold with addition of growth factors, bone xenograft with biomimetic peptides, collagen I composite with bioceramics and a titanium alloy with nanostructured surface. During following in vivo study, we implanted a cell-free scaffold made of PCL, calcium phosphate and IGF-1, bFGF, TGFβ1 and BMP-2 to osteochondral defect. Unfortunately, addition of growth factors resulted in pathological inflammatory process despite clear beneficial effect in vitro. Likewise, the biomimetic peptide sequences promoted osteogenic differentiation of human mesenchymal stem cells. Addition of certain bioceramics influenced the scaffold morphology in the manner of pore size. However, we did not observe any effect of the surface characteristics on cell behavior. The cells were influenced rather by certain material. On the other hand, surface modification of titanium scaffold by anodic oxidation revealed that the most suitable...
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Changes in placental angiogenesis and their impact on fetal intrauterine growth restriction
Kudějová, Alena ; Švandová, Ivana (advisor) ; Magner, Martin (referee)
Any pathological interference with normal vascular development of placenta may have a critical impact on fetal growth and development. The proliferation and differentiation of several cell types play a very important role in the vascular system of placenta. The main factors taking part in the vascular development of placenta include cell elements (e.g. trofoblast, stromal chorion cells, haemangiogenic progenitors), the extracellular matrix, growth factors and cytokines (e.g. VEGF, PlGF, Ang-1,2 and bFGF). The extrinsic factors may also influence the partial oxygen pressure, nutritients availability, and/or the blood perfusion in placenta. Placental ischaemia leading to the worsening of uteroplacentar perfusion is the most common cause of the intrauterine growth retardation (IUGR). The IUGR development is then the result of insufficient prolongation, branching, and dilatation of capillary loops during the formation of terminal villi. Published studies focusing on growth factors in placentas from physiological pregnancies and pregnancies with IUGR do not give clear results. This BSc. Thesis is a review focused on up-to-date-known data concerning changes in placental angiogenesis and their impact on IUGR. Key words: placenta, angiogenesis, IUGR, pregnancy
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Preparation, characterization and testing of blood derivatives for applications in regenerative medicine
Sovková, Věra ; Amler, Evžen (advisor) ; Kolářová, Hana (referee) ; Vránová, Jana (referee)
Platelet products can be used, thanks to the broad range of bioactive molecules, either as a supplement for cell cultering in vitro alone or for development of cell- or cell-free scaffolds in diverse fields in regenerative medicine. The aim of this study was to prepare several types of platelet products. The concentration of selected molecules were observed. Subsequently, these products were tested with cell cultures in vitro alone or in combination with nanofibres scaffolds prepared by electrospinning or centrifugal spinning. It was found out, that platelets products contains chemokine RANTES and growth factor PDGF in the highest concentrations. It was further discovered the content of pro and antiinflammatory in terleukins and other growth factors. Platelet lysat in concentration 7% is sufficient to replace FBS in keratinocytes and fibroblasts cultures. In the other experiments, platelets in different concentrations were adhered to the scaffolds prepared by electrospinning and centrifugal spinning. Thus prepared scaffolds promote the proliferation and viability of all tested cell types in dose-dependent manner. In the last experiment, the individual components of platelet concentrate were separated and characterized. Their effect to the cell culture were tested. It was examinated the synergic...
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Insulin like and other growth factors and tumors
Kučera, Radek ; Topolčan, Ondřej (advisor) ; Klimeš, Jiří (referee) ; Šafarčík, Kristian (referee)
Inzulinu podobné a jiné růstové faktory a nádory Insulin like and other growth factors and tumors Disertační práce Plzeň 2011 Mgr. Radek Kučera Summary The interaction between growth factors and cancer incidence and development has been discussed recently. First works suggesting possible connection between increased production of insulin like growth factors with an increased incidence of tumors came already from early 80s of the last century. The aim of my work was focused on evaluation of possible relation between insulin like growth factors or their binding proteins and tumors from different perspectives, to assess their significance and try to determine what role the IGFs and IGFBPs may play in the current tumor diagnostics. The work itself is divided into two parts, the theoretical and practical ones. In the theoretical part, I summarized the action of IGF in human body and also focused on previous findings on the role of IGF system in cancer diagnoses. In the practical part, divided into three subsections, I focused on investigation of relations in a large set of patients with different cancer diagnoses, in a group of women with breast cancer and, finally, I focused on changes of IGF1 levels during follow-up. Theoretical part Insulin-like growth factors (IGF) are peptides, that participate on growth...
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Insulin like and other growth factors and tumors
Kučera, Radek ; Topolčan, Ondřej (advisor) ; Klimeš, Jiří (referee) ; Šafarčík, Kristián (referee)
Inzulinu podobné a jiné růstové faktory a nádory Insulin like and other growth factors and tumors Disertační práce Plzeň 2011 Mgr. Radek Kučera Summary The interaction between growth factors and cancer incidence and development has been discussed recently. First works suggesting possible connection between increased production of insulin like growth factors with an increased incidence of tumors came already from early 80s of the last century. The aim of my work was focused on evaluation of possible relation between insulin like growth factors or their binding proteins and tumors from different perspectives, to assess their significance and try to determine what role the IGFs and IGFBPs may play in the current tumor diagnostics. The work itself is divided into two parts, the theoretical and practical ones. In the theoretical part, I summarized the action of IGF in human body and also focused on previous findings on the role of IGF system in cancer diagnoses. In the practical part, divided into three subsections, I focused on investigation of relations in a large set of patients with different cancer diagnoses, in a group of women with breast cancer and, finally, I focused on changes of IGF1 levels during follow-up. Theoretical part Insulin-like growth factors (IGF) are peptides, that participate on growth...
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