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Coculture of cells in vitro for bone regeneration
Sloupová, Lenka ; Filová, Eva (advisor) ; Tlapáková, Tereza (referee)
Cocultivation of two (or more) cell types in vitro leading to the formation of functioning bone tissue, later inserted into the damaged area, could be a solution for patients for whom the current methods (e.g. use of human bone grafts) are insufficient. In coculture, one cell type is used to accomplish osteogenesis, while the other is used for angiogenesis, because the limit of diffusion of O2 and essential nutrients is only 200 μm, which means that establishing a vascular network in vitro should prevent the new bone tissue from dying after implantation. Creation and understanding of a functioning coculture in vitro are crucial for developing a coculture successful in vivo. This work summarises and compares information about the influence of in vitro cocultivation on proliferation, osteogenesis and angiogenesis in coculture which uses osteoblasts (or osteoprogenitors), bone marrow mesenchymal stem cells (BMSC) or adipose derived mesenchymal stem cells (ADSC) as it's osteogenic cell type combined with various endothelial cell types. In order to understand the impact of cocultivation on these processes, one chapter deals with interactions between cocultured cell types. Keywords coculture, osteogenesis, angiogenesis, in vitro, osteoblasts, BMSC, ADSC
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Formation of blastema during limb regeneration in Amphibia
Paušlyová, Lucia ; Tlapáková, Tereza (advisor) ; Paňková, Daniela (referee)
Total limb regeneration among vertebrates is basically restricted to some amphibians. Urodeles have the ability to regenerate amputated limbs through their life span. Anurans have the ability of complete regeneration of amputated limbs only in their larval stage. The key process of the limb regeneration is the formation of undifferentiated cell group which is called blastema. There are many cell types that contribute to formation of the blastema while the most important part in this process belongs to the skeleton muscle tissue and dermal fibroblasts. Another critical factor in formation of the blastema and its growth are the nerves in the area of wound and neurotrophic factors produced by them. In the last 20 years it has been great improvement in using different markers for tracking the fate of blastema cells.
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The role of integrins in gametes prior to fertilization and during their interaction
Foldynová, Veronika ; Komrsková, Kateřina (advisor) ; Tlapáková, Tereza (referee)
Integrins are heterodimeric transmembrane glycoproteins that represent a large group of cell adhesion receptors involved in cell-cell, cell-extracellular matrix and cell-pathogen interacti- ons. Up to now, 24 different integrin heterodimers have been detected in mammals. They are involved in a wide range of processes such as immune response, lymphocyte homing, platelet aggregation, also in wound healing, cell differentiation, migration, proliferation and even in cell survival. Integrins have also been detected on germ cells and are now known to play an important role in reproductive processes such as fertilization, embryo implan- tation, and embryonic development. The main aim of this thesis is to introduce integrins from the perspective of reproduction, integrin physiology, occurrence and localization of individual subunits in male and female germ cells. A significant part of the work is devoted to a discussion of the role of integrins in gametes, both during maturation processes (egg maturation in the ovary, capacitation and the acrosomal reaction of sperm), in sperm mi- gration, oviductal reservoir formation and their direct and indirect involvement in adhesion and fusion of the gametes during fertilization. Keywords: integrins, sperm, egg, fusion, fertilization
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Characterization of the porcine induced pluripotent stem cells
Svobodová, Eliška ; Vodička, Petr (advisor) ; Tlapáková, Tereza (referee)
Induced pluripotent stem cells (iPSCs) are the cells established by introducing several transcription factors into the somatic cells and culturing them in embryonal stem cell (ESCs) culture conditions. Factors used for the establishment of the first iPSCs are OCT3/4, SOX2, KLF4 and c-MYC. IPSCs created by these means resemble closely to the ESCs. IPS technology may be used to derive iPS cells of individual pacients and apply these cells for their treatment in the cases where the use of ESCs represents an ethical and imunological problem. Therefore, it is important to establish an appropriate animal model for the longtime safety testing of iPSCs before acceding to their medicinal aplication.
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The Role of Bmp4 and Bmp8b proteins in genesis of mouse primordial germ cells
Vegrichtová, Markéta ; Tlapáková, Tereza (advisor) ; Petelák, Aleš (referee)
Primordial germ cells are precursors of sexual breeding organisms. For induction of their formation are necessary growth factors from extraembryonic ectoderm. There are key proteins BMP4 a BMP8b among these factors. These proteins form both homodimers and heterodimers and through receptor complexes and SMAD proteins are indicating the competent cells of proximal epiblast, which give rise to the primordial germ cells and other structures of embryonic mezoderm. Although BMP4 and BMP8b proteins signal similarly, they are mutually indispensable and though for induction of primordial germ cells are both necessary. Up to now, it was proposed several models of their interaction, but not yet completely accurate.
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Description of Xenopus tropicalis pre-Sertoli cells in tissue culture
Ševčíková Jonáková, Jana ; Tlapáková, Tereza (advisor) ; Drda Morávková, Alena (referee)
5 Abstract Xenopus tropicalis is an important model organism used in developmental and cell biology. Long term primary mixed cell culture, derived from juvenile testis of Xenopus tropicalis, is the first tissue culture of this specie. Characterization of this tissue culture is important for its further handling and subsequent experiments. The culture is composed of Sertoli cell precursors. Feeder layer of this cells enables long term germinal stem cell cultivation. Influence of condicioned medium plays important role during the cell culture cultivation. Result of this task approved effect of condicioned medium for cell culture growth. Cell culture and testis gene expression analyse revealed, that in process of Sertoli cell differentiation many growth and differentiation factors are involved. Presence of marker genes has been approved by RT-PCR and in situ hybridization. This in vitro system can contribute to clarification of different roles of pre-Sertoli and Sertoli cells in testis differentiation, and therefore it establishes a new spermatogenesis model offering the possibility to distinguish parts of the pre-Sertoli and Sertoli cells differentiation, which is difficult in mammal models. Keywords: Xenopus tropicalis, tissue culture, pre-Sertoli cells, Sertoli cells, testicular differentiation, gene expression
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The role of NG2 glycoprotein in cancer cell invasiveness
Obr, Adam ; Brábek, Jan (advisor) ; Tlapáková, Tereza (referee)
NG2 proteglycan is a novel membrane - spanning proteoglycan, expressed in general in developing tissue whose cells are characteristic for its increased level of proliferation and motility. NG2 proteoglycan is considered to be an anchor for cell adhesion capabilities on different substrata as well as a signaling transmembrane structure which is capable of affecting actin cytoskeleton and causing increased cell migration. This bibliographic search shows the considered effect of NG2 proteoglycan to the migration abilities of cancer cells via different molecular mechanisms, such as NG2 - mediated, integrin - independent cell interactions with collagens and other ECM substrata, effect of phosphorylation with two different kinases leading to diverse signaling and different behavior in response to phosphorylation and finally the interaction with scaffolding protein MUPP1 and possible connection with signaling pathway to RhoA GTPase, which is involved in cytoskeleton regulation.
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Potential of stem cell therapy for diabetic retinopathy
Palacká, Kateřina ; Holáň, Vladimír (advisor) ; Tlapáková, Tereza (referee)
Diabetic retinopathy is retinal disease causing irreversible cell damage and consequently a loss of vision. Current treatment protocols have many limitations and are associated with serious site effects. A possible treatment options for retinal degenerative diseases is a use of stem cells. There are different types of stem cells. These include embryonal stem cells, induced pluripotent stem cells and cells from an adult organism, among which we can include mesenchymal stem cells (MSCs). MSCs can be found in almost all tissues of the adult organism. MSCs can migrate to the site of damage, regulate development of inflammation in retina, suppress the formation of fibrovascular scars and replace damaged cells such as nerve cells, photoreceptors and epithelial pigment cells. Application of MSCs could be a promising treatment for degenerative retinal diseases.
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Changes in embryonal programing induced by diabetes mellitus
Landsmann, Lukáš ; Pavlínková, Gabriela (advisor) ; Tlapáková, Tereza (referee)
Embryonic development is sensitive to environmental changes. These changes may lead to changes in the embryonic programming. Changes in programming embryos can occur due to inadequate nutrition, stress, treatment with chemicals and also due to diabetes. Epigenome reacts sensitively to environmental factors regulating gene transcriptional activity. Changes in the epigenome lead to a changes in gene expression, which can have a negative impact on the physiology and metabolism of organism. Maternal diabetes may alter embryonic and fetal development and may result in diabetic embryopathy. Furthermore, maternal diabetic enviromental plays an important role in the predisposition of offspring to a number of chronic diseases later in life. The offspring of diabetic pregnancies demonstrate differences in metabolic, cardiovascular, and inflammatory variables, compared to the offspring of nondiabetic mothers. This thesis summarizes the genetic and epigenetic factors involved in the development of diabetic embryopathy and in the embryonic programming. Key words: Diabetes mellitus, diabetic embryopathy, transcriptional regulation, genetic and epigenetic factors , embryonic programming, genome
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