|
|
Monitoring of mesenchymal stem cell migration in the extracellular matrix
Zumberg, Inna ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
This master’s thesis contains a description of mesenchymal stem cells (MSC). The work includes knowledge about the process of migration and differentiation of MSCs. The theoretical part of this thesis also deals with the problem of cell cultivation in 2D and 3D environments. The most used materials for creating 3D scaffolds are described here. The practical part contains the description of the cell culture protocol for passaging and also describes the experiment in the cell laboratory. The results of the experiment are discussed using confocal microscope images. The proposed experiment was tested with a sufficient number of repetitions. The processing of microscopic images was performed in the MATLAB programming environment.
|
|
|
Study of mesenchymal stem cell migration in the extracellular matrix based on principles of chemotaxis
Scholasterová, Viktorie ; Fohlerová, Zdenka (referee) ; Chmelíková, Larisa (advisor)
This thesis engages in a study of mesenchymal stem cell migration in extracellular matrix based on principles of chemotaxis. First, attention is focused on a theoretical part associated with a clarification of basic terms such as extracellular matrix, migration, confocal microscopy, mesenchymal stem cells or chemotaxis. There is also included a list and a description of some basic methods for monitoring cell migration and a more detailed description of a method called transwell assay, which has been chosen for an experiment in a practical part of this thesis. This part includes protocols of individual steps for the preparation of the experiment, the procedure of data processing obtained by scanning cells with a confocal microscope and a description of the resulting confluence values.
|
|
|
3D models of brain tumors
Fišer, Ondřej ; Novák, Josef (advisor) ; Bohačiaková, Dáša (referee)
Despite intensive research, glioblastoma multiforme remains one of the tumours of the central nervous system with the worst prognosis. The ability of glioblastoma cells to infiltrate brain tissue by forming invasive microtubular structures is stimulated by contact with adjacent non- tumor cells. Intercellular communication and the influence of the extracellular matrix create a specific microenvironment that affects cell signaling, proliferation, differentiation and response to pharmaceuticals. The recurrent form of glioblastoma often displays a much faster progression than the initial disease, which is attributed to the development of resistance to therapeutics and the preservation of the proliferative capacity of some tumour cells. The discovery of the stem- cells ability to self-aggregate in suspension has led to the creation of 3D in vitro models - brain organoids. They are much more complex that the established 2D models and their heterogeneity provides an environment simulating the in vivo state. This thesis aims to describe their use in brain tumour research and techniques for culturing 3D aggregates of neural lineage formed from induced or embryonic human stem cells with respect to their gradually increasing complexity. It also presents methods of addressing issues of hypoxia, organoid...
|
|
|
Proteomic analysis of engineered hyaline cartilage
Kaňovská, Zuzana ; Vičarová,, Petra (referee) ; doc. RNDr. Irena Koutná, Ph.D (advisor)
The aim of this thesis was to observe the expansion of chondrocytes, specialized cartilage cells, in vitro in culture medium containing platelet rich plasma, to evaluate the effect of the addition of growth factors TGF-1 and IGF-1 to the medium and to explore the possibility of proteomic analysis application in the development of cartilage joint replacements. Chondrocytes were isolated form articular hyaline cartilage samples acquired in collaboration with the FN Brno and expanded in media. The resulting constructs were subsequently analyzed by immunocytochemical method, flow cytometry and proteomic analysis using LC-MS/MS.
|
|
|
Regeneration and structural changes of the nerve tissue after the extracellular matrix modification
Vallová, Jana ; Machová Urdzíková, Lucia (advisor) ; Hampl, Aleš (referee) ; Hubálek Kalbáčová, Marie (referee)
Regeneration and structural changes of the nerve tissue after the extracellular matrix modification. Abstract ECM modification may contribute to changes in nerve tissue plasticity. Therefore, 4- methylubulliferone is used in our study, which disrupted the structure of the perineuronal networks surrounding some types of neurons responsible for the formation of memory traces. Mice were fed a 4-MU diet (6.7 mg / g / day) for 6 months, which improved their memory skills in a spontaneous recognition test without a significant adverse effect on the kidneys, liver, or joints.In the next part of the study, an extracellular matrix (UC-ECM) was derived from fetal human umbilical cord tissue also generated as biomimetic hydrogel. Due to the generated UC-ECM's low stability and rapid degradation, the structure was stabilized by covalent genipin bonding. Stabilization with 1 mM genipine increased the biological stability of the material. UC-ECM as well as ECM/G didn't show toxicity in vitro in mesenchymal stem cell proliferation; axonal budding or neural stem cell growth and differentiation were not adversely affected. The biocompatibility of both materials was verified in vivo by applying the material to an intracortical photothrombotic rat lesion, where gelation and infiltration of the lesion and hydrogel by host...
|
|
|
Studies of intercellular interactions in tumours
Jechová, Alžběta ; Smetana, Karel (advisor) ; Skalníková, Helena (referee) ; Masařík, Michal (referee)
Beside tumor cells themselves, tumors consist of many non-malignantly transformed cellular elements and an extracellular matrix. This so-called tumor microenvironment, or stroma, significantly influences the biological properties of the tumor through intercellular interactions. In this thesis I have focused on the study of tumor-associated fibroblasts in squamous cell carcinomas of the head and neck, malignant melanoma and glioblastoma. The data show the presence of cells with mesenchymal characteristics, present even in the glioblastoma stroma, which could potentially have a positive effect on proliferative activity and invasiveness of glioblastoma cells. In malignant melanoma, the presence of keratinocytes should also be considered, as they are the major cells of the epidermis influencing tumor melanocytes. The conditioned medium from UVB irradiated keratinocytes and non-irradiated fibroblasts stimulates the invasion of malignant melanoma cells. Targeting the tumor stroma may be a new direction in oncological therapy, so we have focused on the influence of synthetic polyamine on the formation of myofibroblasts, which are an active part of the population of tumor-associated fibroblasts. The tested polyamine prevents the formation of myofibroblasts but has no effect on those already formed nor on...
|
|
|
The role of ageing in the changes of the brain extracellular matrix and extracellular space properties
Kamenická, Monika ; Vargová, Lýdia (advisor) ; Růžička, Jiří (referee)
The process of aging causes the major changes in nervous tissue such as changes in the size of brain, architecture of glial cells and extracellular matrix. The size of brain is on the decrease as consequence of aging and there is a change of molecules as well as morphology at all levels. Extracellular space (ECS) is interstitium important especially in communication between cells mediated by diffusion. The limit of diffusion in extracellular space is given by size of ECS, which is discribed by volume fraction and tortuosity, that reflect amount of diffusion barriers. The changes of ECS diffusion parameters during aging were measured by real-time iontophoretic method in four parts of brain (cortex - Cx, hippocampus - Hp, inferior colliculus - IC and corpus trapezoideum - TB). Further, we studied influence of deficiency of Bral2 link protein at differences of ECS diffusion parameters and importance of Bral2 protein at aging and regulation mechanisms of cytotoxic brain edema. Our results show, that aging leads to decreasing of ECS volume v Cx and Hp, but it was not observed in IC and TB, where the intact perineuronal nets act like protecting shield against the degenerative disease induced by aging. However, small differences in composition of perineuronal nets, deficiency of Bral2 link protein, may...
|
|
|
Extracellular matrix in yeast populations
Novotná, Pavla ; Kuthan, Martin (advisor) ; Dvořáček, Lukáš (referee)
The microorganisms in a natural environment are frequently found in multicellular forms, most commonly in biofilms. Biofilm is characterized as a community of cells living at the interface of two environments, embedded in the extracellular matrix. ECM is a significant component of biofilms in yeast populations. Extracellular matrix acts as a protective barrier and allows cells to survive under adverse conditions and better compete with other microorganisms. It also forms an effective barrier against antibiotics and other harmful substances, what makes biofilms a serious problem in medicine and industry. Formation of the matrix may be influenced by the morphological forms of colonies. Increased formation of ECM is commonly connected in phenotypic switching in response to changes of their living conditions. The compositeion of the ECM is a genus-and species-specific. The main component of the matrix consists of polysaccharides and proteins.
|
|
|
Mesenchymal stromal cells and biological scaffolds for neural tissue regeneration
Kočí, Zuzana ; Kubinová, Šárka (advisor) ; Filová, Eva (referee) ; Zach, Petr (referee)
Despite tremendous progress in medicine, injuries of the adult central neural system remain without satisfactory solution. Regenerative medicine employs tissue engineering, cellular therapies, medical devices, gene therapy, or growth factors with the aim to bridge the lesion, re-establish lost connections and enhance endogenous repair in order to restore neural function. The aim of my thesis was to evaluate therapeutic potential of two approaches, transplantation of human mesenchymal stromal cells (hMSCs) and biological scaffolds derived from extracellular matrix (ECM) for neural regeneration, particularly in models of spinal cord injury (SCI). First, hMSCs from various sources - bone marrow (BM), adipose tissue (AT) and Wharton's jelly (WJ) - were isolated and characterized in vitro. All cell types met the minimal criteria for MSC phenotype and displayed similar properties in terms of their surface marker expression, differentiation potential, migratory capacity, and secretion of cytokines and growth factors. On the other hand, the cell yield from WJ and AT was significantly higher, and MSCs isolated from these tissues proliferated better than from BM. Therapeutic effect of intrathecal application of hWJ-MSCs was then evaluated in SCI compression model in rats. The effect of low (0.5 million) and...
|
|
|
Functionalized nanofiber system for accelerated regeneration of internal tissues
Kráľovič, Martin ; Amler, Evžen (advisor) ; Gášková, Dana (referee) ; Chvojka, Jiří (referee)
Functionalized nanofiber system for accelerated regeneration of internal tissues Abstract Functionalization can adjust the properties of nanofibers prepared by electrospinning to best replace the missing extracellular matrix in the healing wound. In the rabbit intestinal anastomoses, functionalization of nanofibers by cryogenic fractionation has proven to be crucial. While the use of functionalized PVA and chitosan nanofiber membranes led to intestinal strictures and severe inflammation, further functionalization of PVA and chitosan nanofibers by their cryogenic fractionation led to the possibility of reducing the polymer dose, which significantly reduced the inflammatory response and significantly increased the biocompatibility of the material used. The application of fractionated nanofibers increased the fraction of microvessels, fibroblasts and collagen, which resulted in an increase in the biomechanical strength of healed anastomoses. The functionalization of the polycaprolactone nanofibers by special adhesion to the polypropylene mesh made it possible to use them as a support for the healing of the fascia in the abdominal wall of pigs. Functionalization of PVA nanofibers with glyoxal led to their increased stability in the aqueous environment. By applying a membrane of PVA functionalized nanofibers to...
|
guest ::
