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Selection and validation of reference genes for relative mRNA quantification in human liver slices
Studenovská, Petra ; Boušová, Iva (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Petra Studenovská Supervisor: doc. PharmDr. Boušová Iva, Ph.D. Title of diploma thesis: Selection and validation of reference genes for relative mRNA quantification in human liver slices The use of precision-cut liver slices is often employed in vitro system in the biochemical, pharmacological and toxicological studies. This is mainly due to the preservation of the cellular tissue architecture and therefore its functions. The aim of this diploma thesis was to select and validate reference genes for the relative quantification of mRNA in human liver slices. Liver tissues obtained from 3 patients of different age, diagnosis and pharmacotherapy were used as samples. Liver slices, prepared from samples of liver tissue, were exposed to dimethyl sulfoxide (control) or known inducers of cytochrome P450, rifampicin (10 µM) and β-naphthoflavone (10 µM). Subsequently, the stability of the expression of reference genes was verified using RefFinder program, which integrates the programs of the geNorm, BestKeeper, NormFinder and comparative ΔCT method. The optimal number of reference genes that should be used to evaluate the relative gene expression of target genes was determined using the geNorm program....
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Selection and validation of reference genes for relative mRNA quantification in human liver slices
Studenovská, Petra ; Boušová, Iva (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Petra Studenovská Supervisor: doc. PharmDr. Boušová Iva, Ph.D. Title of diploma thesis: Selection and validation of reference genes for relative mRNA quantification in human liver slices The use of precision-cut liver slices is often employed in vitro system in the biochemical, pharmacological and toxicological studies. This is mainly due to the preservation of the cellular tissue architecture and therefore its functions. The aim of this diploma thesis was to select and validate reference genes for the relative quantification of mRNA in human liver slices. Liver tissues obtained from 3 patients of different age, diagnosis and pharmacotherapy were used as samples. Liver slices, prepared from samples of liver tissue, were exposed to dimethyl sulfoxide (control) or known inducers of cytochrome P450, rifampicin (10 µM) and β-naphthoflavone (10 µM). Subsequently, the stability of the expression of reference genes was verified using RefFinder program, which integrates the programs of the geNorm, BestKeeper, NormFinder and comparative ΔCT method. The optimal number of reference genes that should be used to evaluate the relative gene expression of target genes was determined using the geNorm program....
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