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Preparation of Xenopus tropicalis transgenic testicular stem cell culture.
Vegrichtová, Markéta ; Tlapáková, Tereza (advisor) ; Drobná Krejčí, Eliška (referee)
Testicular stem cells (TSCs) are relatively accessible potential source of pluripotent cells, which are particularly important for their application in regenerative medicine. Xenopus tropicalis is a useful model organism to study the migration and differentiation potential of stem cells. This amphibian is characteristic by outer fecundation and embryonic development of a great amount of embryos after fertilization. Oocytes and embryos are large enough (about 1 mm) to be suitable for micromanipulation micromanipulations. Laboratory of Developmental Biology, Faculty of Science, Charles University in Prague succeeded in the establishment of a mixed cell culture of TSCs growing on feeder layer of pre- Sertoli cells. This culture was derived from the testes of juvenile Xenopus tropicalis male. In the study of their differentiation potential it was found, that leukemia inhibitory factor (LIF) is the decisive factor allowing rapid proliferation of stem cells and their forming into characteristic colonies. This protein is produced by both types of cells which are present in the culture. The mouse LIF has the same positive effect on the proliferative potential of stem cells, which points at the evolutionary conservation of metabolic pathways associated with the maintenance of the stemness. RT-PCR analysis...
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Cell surface CD47 expression in cancer stem cell-targeted tumor therapy
Kuzmík, Ján ; Drbal, Karel (advisor) ; Černý, Jan (referee)
CD47 is a transmembrane glycoprotein with a high expression in both, healthy and cancer (stem) cells. Level of the CD47 expression is negatively correlated with survival of cancer patients. Binding of CD47 to SIRPα, localized on a phagocyte, triggers intracellular signaling cascade. The final effect of this cascade is dephosphorylation of nonmuscle myosin-IIA, which disrupts its function and accumulation to phagocytic synapse. The blockage of CD47-SIRPα signaling pathway in a presence of the pro-phagocytic signal induces phagocytosis of cancer cells. Afterwards, phagocytes can serve as the antigen presenting cells and prime T cell response. Role of CD47-SIRPα signaling pathway in immunity has established this pathway as a target of cancer therapy testing. Preclinical research has identified a positive therapeutic effect of blocking this signaling pathway. Nowadays, the first phase of clinical trials is being conducted. The most prevalent approach of blocking CD47-SIRPα signaling pathway in therapy is the use of anti-CD47 blocking monoclonal antibodies, which cause mild anemia. However, alternative approaches of blocking this pathway are also being developed. In this bachelor thesis, I have summarized the research related to the blockage of CD47-SIRPα signaling pathway as a cancer therapy.
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Preparation of Xenopus tropicalis transgenic testicular stem cell culture.
Vegrichtová, Markéta ; Tlapáková, Tereza (advisor) ; Drobná Krejčí, Eliška (referee)
Testicular stem cells (TSCs) are relatively accessible potential source of pluripotent cells, which are particularly important for their application in regenerative medicine. Xenopus tropicalis is a useful model organism to study the migration and differentiation potential of stem cells. This amphibian is characteristic by outer fecundation and embryonic development of a great amount of embryos after fertilization. Oocytes and embryos are large enough (about 1 mm) to be suitable for micromanipulation micromanipulations. Laboratory of Developmental Biology, Faculty of Science, Charles University in Prague succeeded in the establishment of a mixed cell culture of TSCs growing on feeder layer of pre- Sertoli cells. This culture was derived from the testes of juvenile Xenopus tropicalis male. In the study of their differentiation potential it was found, that leukemia inhibitory factor (LIF) is the decisive factor allowing rapid proliferation of stem cells and their forming into characteristic colonies. This protein is produced by both types of cells which are present in the culture. The mouse LIF has the same positive effect on the proliferative potential of stem cells, which points at the evolutionary conservation of metabolic pathways associated with the maintenance of the stemness. RT-PCR analysis...
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