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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
Optimization of miRNA analysis in fine-needle biopsy samples of pancreatic cancer tissue.
 Čuperková, Romana ; Benešová, Lucie (advisor) ; Kuthan, Martin (referee)
Pancreatic cancer (PC) is extremely severe malignant disease with a five-year survival of less than 5%. Currently there is no reliable tool for the diagnosis of PC in its early stages. At the time of clinical symptoms most patients are in an advanced stage of the disease and the treatment does not usually have a significant effect. For these reasons emphasis is gradually shifting to the search for the suitable molecular markers for improvement of the diagnosis and assessment of the survival prognosis with respect to a possibility of surgical treatment. MiRNA represent one of the most promising markers, although, their examination in pancreatic tissue is a complicated process. One of the reasons is the very small amount of the source material coming from a fine needle biopsy. A second cause of problems is the subtle character of the pancreatic tissue resulting in significantly lower yields of molecular genetic analysis when compared to other epithelial tissues. An additional negative factor is heterogeneity of the tissue resulting in disproportionate representation of tumor cells within the sample. A suitable choice of procedures for isolation of nucleic acids (NA) and subsequent analysis including quantification of tumor cells is critical for accurate evaluation of the miRNA levels. This work is...
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Optimization of Culture Medium for HEK293 Cell Line
 Čuperková, Romana ; Vaněk, Ondřej (advisor) ; Šácha, Pavel (referee)
HEK293 is a human cell line derived from embryonic kidney cells and is a frequently used system for the production of recombinant proteins. This work dealt with optimization of the composition of serum-free medium for HEK293S and HEK293T cell lines as a compensation for expensive commercial media. The growth of culture and expression of reporter proteins SEAP and GFP was monitored as the markers. I managed to create a new medium which contained, among other compounds, insulin (1 mg/l), transferrin (5 mg/l) and a mixture of trace elements. During the cultivation in a mixture of commercial medium EX CELL 293 with my new medium 293S cells grew faster than during the cultivation in commercial media (doubling time 20,47 ± 2,68 hours (srel = 13,1 %)). It seems that the new medium is suitable for transfection of HEK293 cell lines with a relatively high expression of recombinant proteins. Transfection ratio of DNA:PEI (w/w) for this medium is 1:2 to 1:3.
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