|
|
Mechanisms determining the identity of intestinal epithelial cells
Berková, Linda ; Valenta, Tomáš (advisor) ; Krylov, Vladimír (referee) ; Klíma, Jiří (referee)
(ENG) The intestinal epithelium has been heavily studied for decades. It is a traditional model to study the natural renewal of tissue during homeostasis and upon the damage, carcinoma establishment and progression and regulation of stem cells by Wnt signalling. Since the main interest in the tissue is focused on epithelial stem cells, it has been described quite recently that also all differentiated epithelial cells from the small intestine including enterocytes undergo drastic changes in properties during their short life. This phenomenon came to be known as villus zonation. In parallel with that observation, the heterogeneity of intestinal mesenchymal cells and their impact on intestinal epithelium have been gradually revealed. Distinct populations of mesenchymal cells secrete specific signalling molecules, which determine the destiny of epithelial cells. Those signalling molecules regulate stem cell maintenance as well as villus zonation. Our work combines those novel findings of intestinal enterocytes and mesenchymal cells. We studied the effect of distinct Bmp ligands produced by mesenchymal cells on the differentiation of enterocytes. We treated small intestinal epithelial cells with recombinant Bmp proteins in vitro and analysed changes in their gene expression by RNA sequencing. We...
|
|
|
Generation and application of induced pluripotent stem cells in hematology
Berková, Linda ; Láníková, Lucie (advisor) ; Krulová, Magdaléna (referee)
Induced pluripotent stem cells (iPSC) are one of the key discoveries in cell biology of the last decade. These cells are pluripotent stem cells derived from differentiated somatic cells while having used only four exogenous transcription factors. Pluripotent cells, which can be derived from somatic cells carrying genetic mutation, have a great potential to be used in the testing of new drugs and in discovering molecular mechanisms of genetic disorders. iPSC derived from healthy cells can be used in regenerative medicine. Originally, retroviral vectors were used for delivering reprogramming transcription factors to cells. However such approach is not safe for medicinal use, because of the ability of retroviruses to integrate into the host genome. This fact initiated development of safer delivering methods of transcription factors into the cells. In this work I present the overview of methods which have been used for reprogramming including the most common techniques used to test pluripotency. In addition, I will describe iPSC application options for therapy of genetically determined hematological disorders (sickle cell anemia, β-thalassemia, X-linked chronic granulomatous disease) and for modelling of their molecular mechanism (polycythemia vera). Key words: iPSC, reprogramming, hematological...
|
|
|
Molecular mechanism of autosomal dominant polycythemia
Berková, Linda ; Láníková, Lucie (advisor) ; Kapráľová, Katarína (referee)
All red blood cells, erythrocytes, originate in bone marrow. The process of their differentiation and maturation is called erythopoiesis and is regulated through hormone erythropoietin (EPO), which functions as a stimulatory factor for erythropoiesis. EPO is produced in kidney and its production is regulated by oxygen supplementation. EPO is transported to bone marrow via blood vessels. Chronic overproduction of erythrocytes leads to disease called polycythemia. Polycythemia may be diagnosed for example by measurement of haematocrit or haemoglobin concentration in blood. EPO level may or may not be increased. Patients suffering from polycythemia may or may not have any symptoms. It depends on manifestation level of the disease. The most common symptoms are higher blood pressure, headaches, dizziness, swelling and epistaxis. Recently, the most common treatment of polycythemia is phlebothomy. The aim of this master thesis is to unravel the role of a newly described mutation, which was found among members of one family suffering from polycythemia with increased EPO level. It is single substitution mutation -136 G > A in 5' UTR region of EPO gene. The clones of EPO producing cell lines bearing this mutation were prepared using CRISPR/Cas9 technology. Several experiments performed not only on those cell...
|
|
|
Generation and application of induced pluripotent stem cells in hematology
Berková, Linda ; Láníková, Lucie (advisor) ; Krulová, Magdaléna (referee)
Induced pluripotent stem cells (iPSC) are one of the key discoveries in cell biology of the last decade. These cells are pluripotent stem cells derived from differentiated somatic cells while having used only four exogenous transcription factors. Pluripotent cells, which can be derived from somatic cells carrying genetic mutation, have a great potential to be used in the testing of new drugs and in discovering molecular mechanisms of genetic disorders. iPSC derived from healthy cells can be used in regenerative medicine. Originally, retroviral vectors were used for delivering reprogramming transcription factors to cells. However such approach is not safe for medicinal use, because of the ability of retroviruses to integrate into the host genome. This fact initiated development of safer delivering methods of transcription factors into the cells. In this work I present the overview of methods which have been used for reprogramming including the most common techniques used to test pluripotency. In addition, I will describe iPSC application options for therapy of genetically determined hematological disorders (sickle cell anemia, β-thalassemia, X-linked chronic granulomatous disease) and for modelling of their molecular mechanism (polycythemia vera). Key words: iPSC, reprogramming, hematological...
|
guest ::
RSS feed.