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Human lymphopoiesis and its examination via single-cell analysis
Novák, David ; Kalina, Tomáš (advisor) ; Drbal, Karel (referee)
Development of human B-lymphocytes is a convoluted process. A self-renewing stem cell progenitor in a primary lymphoid tissue commits to the lymphoid lineage. Subsequent B-lineage commitment entails somatic gene recombination processes which lead to the eventual expression of a surface antigen receptor. Functionality of the B-cell receptor, as well as successful testing for autoreactivity by the cell, are preconditions for the differenti- ation of a mature B-lymphocyte. Processes within this development are often investigated using single-cell analysis via flow cytometry, fluorescence-activated cell sorting and mass cytometry. Coupling these high-throughput methods with modern approaches to data analysis carries enormous potential in revealing rare cell populations and aberrant events in haematopoiesis. Keywords: B-lymphocyte, lymphopoiesis, flow cytometry, FACS, mass cytometry, clus- ter analysis, FlowSOM, PCA, t-SNE, Wanderlust.
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Human lymphopoiesis and its examination via single-cell analysis
Novák, David ; Kalina, Tomáš (advisor) ; Drbal, Karel (referee)
Development of human B-lymphocytes is a convoluted process. A self-renewing stem cell progenitor in a primary lymphoid tissue commits to the lymphoid lineage. Subsequent B-lineage commitment entails somatic gene recombination processes which lead to the eventual expression of a surface antigen receptor. Functionality of the B-cell receptor, as well as successful testing for autoreactivity by the cell, are preconditions for the differenti- ation of a mature B-lymphocyte. Processes within this development are often investigated using single-cell analysis via flow cytometry, fluorescence-activated cell sorting and mass cytometry. Coupling these high-throughput methods with modern approaches to data analysis carries enormous potential in revealing rare cell populations and aberrant events in haematopoiesis. Keywords: B-lymphocyte, lymphopoiesis, flow cytometry, FACS, mass cytometry, clus- ter analysis, FlowSOM, PCA, t-SNE, Wanderlust.
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Effect of sperm ubiquitination on the early embryonic development in pig
Kroumanová, Kristýna ; Krylov, Vladimír (advisor) ; Nevoral, Jan (referee)
The ubiquitin-proteasomal system which is the major pathway for intracellular protein degradation is also involved in sperm quality control in the mammalian epididymis. Defective sperm become surface- ubiquitinated during epididymal passage. The level of sperm surface ubiquitination negatively correlates with their quality. Hypothetically it is possible that after fertilization, highly ubiquitinated sperm, naturally present in mammalian ejaculates, would be actively recognized by oocyte (probably via 26S proteasomal complex). Subsequent partial or total sperm degradation should negatively affect the development of the potentially defective embryo. In this study, we examined the effect of sperm ubiquitination on the early embryonic development in pig (Sus scrofa f. domestica) using the method of intracytoplasmic sperm injection (ICSI). In vitro embryonic development to the blastocyst stage after ICSI was comparable with other laboratories. In this study, no significant difference was observed in the formation of pronuclei between oocytes fertilized by lower and highly ubiquitinated sperm cells. On the other hand, significantly better embryonic development to the blastocyst stage was observed in oocytes fertilized by sperm with lower surface ubiquitination (17 %) compared with oocytes fertilized by highly...
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