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Immunomodulatory properties of mesenchymal stem cells - use in therapy
Pavlíková, Michaela ; Stříž, Ilja (referee) ; Krulová, Magdaléna (advisor)
Mesenchymal stem cells (MSC) are extensively studied mainly due to their feasible clinical application. Therapeutic potential of MSC consists not only of the ability to differentiate into mesenchymal cells, ectodermal and endodermal cell lines, but primarily in their immunomodulatory functions. Due to their effect on immune cells, MSC promote the shift of the inflammatory immune response to antiinflammatory. The ability to suppress inflammation, together with their differentiation potential and antiapoptotic potential on the surrounding cells makes MSC a promising tool for treating serious diseases. This work discusses the effect of MSC on the individual cells of the immune system. It focuses on the description of the effect of MSC in four model cases. These are an experimental autoimmune encephalomyelitis, myocardial infarction, diabetes mellitus and skin graft transplantation. The knowledge of the mechanisms of the interactions between MSC and the cells of the immune system, together with the understanding the effect of specific conditions on MSC is essential for their use in clinical therapy. Keywords: mesenchymal stem cells, immunomodulation, autoimmune diseases, transplantation
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Study of pheonotype of the H9c2 cells derived from rat cardiac myoblasts
Kutichová, Martina ; Šimůnek, Tomáš (advisor) ; Vostatková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Martina Kutichová Supervisor: Doc. PharmDr. Tomáš Šimůnek, Ph.D. Supervisor - specialist: Doc. MUDr. Michaela Adamcová, Ph.D. Title of diploma thesis: Study of phenotype H9c2 cells derived from rat cardiac myoblasts The cell-based in vitro investigation represents an important and frequently employed platform in current biomedical research. In experimental cardiovascular research, enzymatically dispersed neonatal or adult cardiomyocytes from various animal species are considered as a "gold standard" for such experiments. However, besides isolated primary cultures, permanent cell lines are becoming extremely popular among cardiovascular scientists. H9c2(2-1) is the first permanent cardiomyocyte-derived cell line. Although these cells are known since the mid-seventies, there is not much information about their phenotype. This study followed the results of the shotgun proteomic analysis, which identified about 1500 proteins. The aim of this thesis was to continue with obtained proteomic results with focus on detection of specific proteins by Western blot method. H9c2 cell were studied for the presence of several muscle and/or cardiac specific proteins in both non-proliferating H9c2...
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Improvement of retroviral vectors for efficient gene transfer
Šenigl, Filip ; Hejnar, Jiří (advisor) ; Šmahel, Michal (referee) ; Ruml, Tomáš (referee)
44 CONCLUSIONS Retroviral vectors are transcriptionally unstable in mammalian cells. The ASLV- derived vectors are the most affected by silencing. We modified the vector by insertion of the CpG island inner element (IE) into the vector LTR in three different positions in either sense or antisense orientation. Each vector variant exhibited a certain extent of stabilization. The vector with insertion of a tandem of two IEs between the enhancer and the promoter was the most stable and exhibited almost no silencing after four months of cultivation in rodent and human cells. The IE comprises two high-affinity Sp1 binding sites. The presence of Sp1 binding sites is important for the protective effect of IE, but at least a part of the entire anti-silencing capacity is maintained in IE with mutated Sp1 sites. We identified the Tvc receptor of ASLV. The tvc gene encodes a 488-amino-acid protein most closely related to mammalian butyrophilins, members of the immunoglobulin protein family. To confirm the identification of the Tvc receptor, we disrupted both tvc alleles in normally susceptible DT40 cells. The DT40 tvc-/- clone was resistant to the ASLV(C) infection. We identified the mutation that results in decreased susceptibility to infection by ASLV subgroups B and D and resistance to ASLV subgroup E of line M...
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The impact of selected sesquiterpenes on the activity of antioxidant enzymes in the differentiated Caco-2 cell line
Verešová, Erika ; Svobodová, Hana (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Erika Verešová Supervisor: PharmDr. Hana Bártíková, Ph.D. Title of thesis: The impact of selected sesquiterpenes on the activity of antioxidant enzymes in the differentiated Caco-2 cell line The cases of cancer diagnosis rapidly continue to grow across the population. It is a modern trend to buy more and more often a variety of herbal food supplements in order to strengthen or restore human health. Many scientists have been researching Myrica rubra plant for its potential therapeutic effects. In our research we used sesquiterpenes isolated from this plant, in order to determine their effects on the activity levels of antioxidant enzymes in a differentiated Caco-2 cell line derived from intestinal epithelium tumor that have maintained characteristics of healthy cells. Caco-2 cells are being used as a basis for testing of substances and their effects on non-cancerous cells. The cells were incubated for 24 and 48 hours with a concentration 25 µg/ml of tested substances. Essential oil from Myrica rubra and selected individual sesquiterpenes (α- humulene, β-caryophyllene and trans-nerolidol) were chosen to be tested. Of the antioxidant enzymes, we determined the enzymes glutathione-S-transferase,...
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SMV-2014-27: Optical system for generation of stable optical frequency
Hrabina, Jan ; Číp, Ondřej ; Čížek, Martin ; Hucl, Václav
The work was oriented to design and development of a stable optical frequency laser standard based on ultra-low noise DFB laser. The laser frequency was locked to selected rotational-vibrational transition of isotopic acetylene gas by the method of linear absorption spectroscopy. The research included: design, investigation and development of narrow-linewidth DFB laser source including of development of ultra-low noise power-supply and control electronics; development of an optical setup equipped with opto-fiber output for automatic locking of the laser to selected transition of the ultra-pure acetylene gas, special custom-design absorption cell development, filled with isotopic acetylene, with optical windows equipped with custom-design antireflection coatings; development of control electronics for automatic searching of absorption lines and automatic locking of the laser frequency to this transition; development of control software.
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Polymeric nanofibrous scaffolds reinforced with diamond and ceramic nanoparticles for bone tissue engineering
Bačáková, Lucie ; Pařízek, Martin ; Staňková, Ľubica ; Novotná, Katarína ; Douglas, T.E.L. ; Brady, M. A. ; Kromka, Alexander ; Potocký, Štěpán ; Stránská, D.
Three types of nanofibrous scaffolds were prepared by electrospining: (1) poly(lactide-co-glycoside) (PLGA) scaffolds reinforced with 23 wt.% of diamond nanoparticles (DNPs), (2) poly(L-lactide) (PLLA) scaffolds with DNPs in concentration ranging from from 0.4 wt.% to 12.3 wt.%, and (3) PLLA scaffolds with 5 wt.% or 15 wt.% of hydroxyapatite (HAp) nanoparticles. The diameter of the nanofibers ranged between 160 and 729 nm. The nanofibers with nanoparticles were thicker and the void spaces among them were smaller. Mechanical properties of the nanoparticle-loaded scaffolds were better, as demonstrated by a rupture test in scaffolds with DNPs and by a creep behavior test in scaffolds with HAp. On PLGA scaffolds with DNPs, the human osteoblast-like MG-63 cells adhered in similar numbers and grew with similar kinetics as on pure PLGA scaffolds. Human bone marrow mesenchymal stem cells grew faster and reached higher population densities on PLGA-DNP scaffolds. However, on PLLA-based scaffolds, the activity of mitochondrial enzymes and concentration of osteocalcin in MG-63 cells decreased with increasing DNP concentration. On the other hand, the metabolic activity of MG-63 cells and content of osteocalcin in these cells were positively correlated with the HAp concentration in PLLA scaffolds. Thus, PLGA nanofibers with 23 wt% of DNPs and PLLA nanofibers with 5 and particularly 15 wt.% of HAp seem to be promising for bone tissue engineering.
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Evolution of algorithm for artificial creatures
Kohout, Jan ; Mráz, František (referee) ; Holan, Tomáš (advisor)
The task of this work is to create an application which allows performing of experiments with evolution of artificial creatures. Behavior of each creature is managed by its algorithm. The algorithm is able to use creature's memory cells (which represent logical variables) and is the genome of the creature. By the crossing-over and mutation, the algorithm evolves. The second aim of this work is to make some tests with the application and describe their results.
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