|
| |
|
|
Preparation of X. tropicalis recombinant growth factors and their characterization in testicular tissue culture.
Borecká, Marianna ; Krylov, Vladimír (advisor) ; Drobná Krejčí, Eliška (referee)
In our Laboratory of Developmental Biology there was established a long term culture derived from Xenopus tropicalis testes. It contains pre-Sertoli cells mostly. They compose a feeder layer allowing cultivation of stem cells, revealing the morphology of spermatogonial stem cells. This diploma thesis was focused on a preparation of two growth factors, FGF2 (fibroblast growth factor 2) and GDNF (glial cell line-derived neurotrophic factor), with the subsequent characterization of their influence at cell culture mentioned above. Factors were selected on the basis of the microenvironmental niche theory, according which FGF2 and GDNF are the most important factors for spermatogonial stem cells proliferation and self-renewal. FGF2 recombinant factor was gained using the expression plasmid pET-15b. Its characterization in the testicular culture brought surprising result. Even a low concentration of FGF2 factor (2.5ng/ml) caused cell detaching and dying. Similar result was previously shown in differentiating osteoblast culture only. More experiments need to be done to prove if apoptose take place and why do testicular cells act this way. Key words: Xenopus tropicalis, FGF2, GDNF, SSC, pre-Seroli cells
|
|
|
Role of genetic factors responsible for development of pancreatic cancer
Borecká, Marianna
Cancer is a major health problem, worldwide, and is the second-most frequent cause of death (www.uzis.cz). Research is urgently necessary to reduce cancer incidence and the costs associated with cancer management, to develop more efficient and effective risk prediction strategies and personalised patient treatment. Germinal mutations in genes that predispose individuals to hereditary cancer syndromes are clinically important and can be used to classify carriers being at high risk of cancer development. Identification of these mutations can influence the prognosis and treatment of probands and can be used to include their family members into high-risk groups with corresponding preventive strategies. This study is focused on the currently underestimated description of newly identified genetic factors among the Czech population that predispose individuals to develop pancreatic ductal adenocarcinoma (PDAC). In 2017, the incidence of PDAC in the Czech Republic was 21 cases per 100,000 persons, and PDAC was the fourth-most frequent cause of death among all cancer diseases (www.svod.cz). Using a variety of screening techniques, which included high-resolution melting (HRM) analysis, Sanger sequencing of whole genes, and next-generation sequencing (NGS), with a CZECANCA panel that was generated in our...
|
|
|
Role of genetic factors responsible for development of pancreatic cancer
Borecká, Marianna ; Janatová, Markéta (advisor) ; Václavíková, Radka (referee) ; Živný, Jan (referee)
Cancer is a major health problem, worldwide, and is the second-most frequent cause of death (www.uzis.cz). Research is urgently necessary to reduce cancer incidence and the costs associated with cancer management, to develop more efficient and effective risk prediction strategies and personalised patient treatment. Germinal mutations in genes that predispose individuals to hereditary cancer syndromes are clinically important and can be used to classify carriers being at high risk of cancer development. Identification of these mutations can influence the prognosis and treatment of probands and can be used to include their family members into high-risk groups with corresponding preventive strategies. This study is focused on the currently underestimated description of newly identified genetic factors among the Czech population that predispose individuals to develop pancreatic ductal adenocarcinoma (PDAC). In 2017, the incidence of PDAC in the Czech Republic was 21 cases per 100,000 persons, and PDAC was the fourth-most frequent cause of death among all cancer diseases (www.svod.cz). Using a variety of screening techniques, which included high-resolution melting (HRM) analysis, Sanger sequencing of whole genes, and next-generation sequencing (NGS), with a CZECANCA panel that was generated in our...
|
|
|
Preparation of X. tropicalis recombinant growth factors and their characterization in testicular tissue culture.
Borecká, Marianna ; Krylov, Vladimír (advisor) ; Drobná Krejčí, Eliška (referee)
In our Laboratory of Developmental Biology there was established a long term culture derived from Xenopus tropicalis testes. It contains pre-Sertoli cells mostly. They compose a feeder layer allowing cultivation of stem cells, revealing the morphology of spermatogonial stem cells. This diploma thesis was focused on a preparation of two growth factors, FGF2 (fibroblast growth factor 2) and GDNF (glial cell line-derived neurotrophic factor), with the subsequent characterization of their influence at cell culture mentioned above. Factors were selected on the basis of the microenvironmental niche theory, according which FGF2 and GDNF are the most important factors for spermatogonial stem cells proliferation and self-renewal. FGF2 recombinant factor was gained using the expression plasmid pET-15b. Its characterization in the testicular culture brought surprising result. Even a low concentration of FGF2 factor (2.5ng/ml) caused cell detaching and dying. Similar result was previously shown in differentiating osteoblast culture only. More experiments need to be done to prove if apoptose take place and why do testicular cells act this way. Key words: Xenopus tropicalis, FGF2, GDNF, SSC, pre-Seroli cells
|
|
| |
guest ::
RSS feed.