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Study of differential potential of spermatogonial stem cells via transplantation in vertebrates
Kodedová, Barbora ; Krylov, Vladimír (advisor) ; Pšenička, Martin (referee)
Spermatogonia, or spermatogonial stem cells are necessary to maintain male fertility. In the complex process of ongoing spermatogenesis in the testes these pluripotent stem cells proliferate and differentiate into sperm cells. In 1994 the first spermatogonial transplantation technique was described in rodents to allow the study of male germ cells. The following series of studies of intra- and inter-species transmission of testicular tissue revealed the regenerative capacity of transplanted spermatogonial stem cells and their possible usage. Recently, spermatogonia transplantation systems are developed in many vertebrates making it possible to study the development of sperm as well as artificial production of male and female gametes derived from germ cell donors. The differentiation potential of spermatogonial stem cells enabled the creation of transgenic organisms by genetic manipulation of isolated spermatogonia and subsequent transplantation into a suitable recipient. Spermatogonial transplantation in the future may find their application in regenerative medicine, the treatment of disorders of spermatogenesis, or serve to preservation of genetic stock of endangered species.
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Cadaveric bone marrow transplantation: effects of hypoxia and metabolic starvation on mouse hematopoietic stem cells
Linhartová, Jana ; Šefc, Luděk (advisor) ; Průcha, Miroslav (referee) ; Šinkorová, Zuzana (referee)
Objectives: Hematopoietic stem cell transplantation (HSCT) is a widely used method for treatment of hematological disorders and some other diseases. However, sometimes a suitable donor of hematopoietic stem cells (HSC) is not found for a patient. Because HSC have been described as cells with low proliferative and metabolic activity, their tolerance to the lack of oxygen or metabolic substrates may be assumed. In this study, we explored cadaveric bone marrow as an alternative source of HSC for HSCT, using a mouse experimental model. In addition, the effect of in vitro metabolic inhibition and short-term in vitro storage (1 - 4 days) on functional properties of mouse HSC was investigated. Methods: C57Bl/6 mice (wild-type or p53-/- ) were used in the experiments. To explore cadaveric HSC, bone marrow (BM) was left in intact femurs at 37řC, 20řC and 4řC under the conditions of ischemia. The bone marrow cells were harvested after defined time periods ranging 0 - 48 hours. For metabolic inhibition, the electron transport chain inhibitor potassium cyanide (KCN) and inhibitor of glycolysis 2-deoxy-D-glucose (2-DG) were used in vitro. To determine the impact of ischemia, metabolic inhibition, or in vitro storage on transplantability of HSC, the competitive repopulation assay using Ly5.1/Ly5.2 congenic model...
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Diagnostic contribution to the evaluation of intervention models in the treatment of type 1 diabetes
Zacharovová, Klára ; Saudek, František (advisor) ; Plecitá, Lydie (referee) ; Mandys, Václav (referee)
During treatment of diabetes mellitus by immunointervention or transplantation, it is necessary to monitor the markers of immune destruction or rejection of surviving insulin producing cells. An aim of this thesis is to improve the possibilities of following autoimmunity and to detect the survival of transplanted pancreatic islet in vivo. Partial aims included vitality testing of isolated islets for transplantation by measurement of respiration activity, observing the process of in vitro labeling of isolated islets with superparamagnetic iron oxide (SPIO) contrast agent for subsequent magnetic resonance imaging (MRI) of islets and observing SPIO particles transport after transplantation. We also studied a new dual paramagnetic contrast agent combined with fluorescein intended for identification of the MRI contrast agent in samples for histology. Further, we assessed autoimmune reaction by evaluation of cytokine response to specific stimulation with auto-antigens. We tried to affect beta-cells destruction by polyclonal anti- thymocyte antibodies in a mouse experimental model. A new method of the islet respiration measurement correlated with other methods of islet quality testing and it was suggested as a diagnostic test before clinical transplantation. Results obtained studying the intercellular...
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Histomorphological Changes in Normal and Pathological Cartilage Tissues and after their Experimental and Clinical Transplantation
Kaňa, Radim ; Nečas, Emanuel (advisor) ; Slípka, Jaroslav (referee) ; Ludvíková, Marie (referee)
1 Abstract Introduction Autologous transplants of the cartilage tissue from the pinna is commonly used in reconstructive surgery of the nasal skeleton. The present study used animal models to elucidate responses of the auricular cartilage to its damage or transplantation to ectopic sites. Histomorphological analysis of changes observed in auricular cartilage including immunohistochemical study of different isoforms of actin and S-100 proteins was performed. Human articular cartilage prepared by in vitro cultivation using artificial scaffolds was also studied after its transplantation. Aims of the study The aim was to study histological changes and expression of chondrocytic markers (α- SMA and S-100 proteins) in intact, artificially traumatised, or in a human auricular cartilage cultivated in culture medium. An attempt to grow human auricular cartilage chondrocytes implanted in vitro into various types of three dimensional scaffolds aimed at testing chondrocyte survival and phenotype both in the culture and after transplantation to immunodeficient mice. A human auricular cartilage transplanted into the nasal skeleton of patients during a reconstruction surgery should be submitted to a histomorphological examination. Research assumed also comparison of the auricular cartilage responses to a damage,...
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The role of IL-17 in kidney transplantation
Menšíková, Markéta ; Krulová, Magdaléna (referee) ; Stříž, Ilja (advisor)
The role of IL-17 in kidney transplantation - abstract Naive CD4+ T-lymphocytes (Thp) can develop into Th17 line in the presence of TGF- and IL-6. Th17 cells are characterized by expression of Ror- t and by production of interleukin-17 (IL-17). It is secreted as a glycoprotein homodimer. Binding to IL-17 receptor (IL-17R), which is present in all cell types, stimulates the production of proinflammatory cytokines and chemokines. The ratio of Th17: Treg in the graft showing signs of rejection is higher than in the graft without rejection. The presence of IL-17 in a culture of proximal tubular epithelial cells (PTEC) stimulates the production of IL-6, IL-8, MCP-1 and C3 complement component. Simultaneous action of IL-17 and CD40L synergistically increases the production of IL-6, IL-8 and RANTES. Signaling from the receptor on the surface of PTEC associated with its increased expression is effected via the src kinase and MAP kinase, and probably leads to the transcription factor NF- B. In rat models of transplantation, the IL-17 appears in allografts on the second day after surgery, the level rises until the fifth day, then decreases and disappears before the death of the animal. IL-17 is not detectable in isografts and negative controls. It appears before the IFN- , which had been considered a trigger of...
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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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Medical Specialist Awareness and Attitude about Organ Donation
Hajšlová, Petra ; Hošťálková, Monika (advisor) ; Vaňková, Milena (referee)
Bachelor thesis is focused on finding the awareness and attitudes of nurses towards organ donation. The work is divided into two parts ̶ theoretical and empirical. In the theoretical part summarizes available data on the history and development in this field, applicable legislation is mentioned, care for organ donors and overview of the current situation of organ donation, along with current trends. Empirical part took the form of quantitative research, which was conducted by anonymous questionnaires. The aim was to determine nurses' awareness about organ donation organization, determine their orientation in the field of diagnosis of brain death, to map the contemporary practice in the care of a dead donor organs and detect personal attitude on the issue of donation. Respondents were nurses that during their practice on the wards encounter this issue. Questionnaires were distributed on anesthesiology and resuscitation department of Central Military Hospital ̶ Military University Hospital in Prague, then to the operating room and intensive inpatient department of intensive and resuscitation care at the Institute for Clinical and Experimental Medicine. Investigation was attended by 96 nurses. In results, we found that the principle of presumed consent to organ donation in the Czech Republic is...
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Opinions of the students of the South Bohemia university about selected ethical problems of the health care
RAZIMOVÁ, Pavlína
This Bachelor thesis is focused on ethical issues in medicine, namely in communication between physician and patient, further on euthanasia, interruption and transplantation. This work is divided into two parts. The theoretical part introduces the theme of the thesis. It provides the basic information about ethics, about the relationship between physician and patient and their communication, about patient´s rights, about euthanasia, interruption and transplantation, too. The practical part reflects opinions of the students of the South Bohemia university in České Budějovice of these ethical matters. The analysis is based on quantitative research, which was performed by anonymous questionnaire in electronic form. The obtained data are processed in tabular and graphic forms.
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Opinions of the adult population of the South Bohemia region about selected ethical problems of the health care
BICANOVÁ, Anna
The theme of this thesis are opinions of adult population of the South Bohemia region about selected ethical problems. These problems involve doctor's relationship with patient, opinions of euthanasia, abortion and transplantation organs and tissues. In the theoretical part are explained and escribed the basic concepts related to this thesis. Furthermore, the theoretical part deals with the views from professional literature on these selected topics. The aim of the thesis is to determine the views of the adult population of South Bohemia on selected ethical issues in health care. The main objectives are the views on the relationships of the doctors with the patients, the introduction of euthanasia in the Czech Republic, a positive or negative view of abortion and opinion on supposed consent to postmorten donating organs. In the practical part the thesis deals with the quantitative research and evaluation of the research questionnaire.
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