Essay about The Cancer Cells And The Body Of Zebrafish Larvae

Essay about The Cancer Cells And The Body Of Zebrafish Larvae

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subsection{Xenotransplantation}

The cancer cells were xenotransplanted into optic tectum, heart and the otic vesicle of zebrafish at 4 dpf (figure ef{fig:injection1}).
Transplantation was done using an Eppendorf Femtojet 4i under a Nikon SMZ1500 macroscope at 1X. Needles were prepared from thin-wall single-barrel glass capillaries (WPI, Hitchin, UK) with a needle puller P-97 Flaming/Brown type micropipette puller (Sutter Instruments, USA) with parameters pull = 230, velocity = 50, time = 125, heat = 650. Removal of the needle tip was dependant on cell density and size, avoiding high injection pressures, as well as injection of fluid only to circumvent rupture of brain compartments.

egin{figure}

centerline{includegraphics[scale=0.11]{tikvi.png}}
caption{ extbf{ The human tumour cell lines were xenotransplanted in three sites in the body of zebrafish larvae.} A) Dorsal view of the head in larval zebrafish at 4 dpf: the orange circle shows the area of the right optic tectum. B) Lateral view of head in larval zebrafish at 4 dpf: the top orange circle shows the otic vesicle and the bottom orange circle surrounds the heart. }
label{fig:injection1}
end{figure}





subsection{Immunosuppression and larvae fixing}

Immunosuppression was introduced at day five post fertilisation in one group of zebrafish with xenotransplanted GBM cells and one no procedure control group. As an immunosuppressive drug 100 $mu$g/mL Dexamethasone (Sigma-Aldrich, Dorset, UK) was used to suppress the development and maturation of T-cells, following the experimental protocol (Langenau et al., 2004). Dexamethasone powder (5 $mu$g) was weighted and dissolved in 1 mL 0.4$\%$ ethanol solution that was...


... middle of paper ...


...(volume) at time t, $alpha$ is a constant describing tumour growth, $eta$ is a constant describing loss of tumour mass as a result of cell death.
subsection{Statistical methods}

Unless stated otherwise, the statistics software used is Minitab 17.

Stem-and-leaf plots and the Anderson-Darling statistic were used to test for normality of data. Data transformations were made using the Johnson transformation function when required. Statistical techniques included box plots, two-tailed Student t-test, and the Kaplan-Meier survival function, evaluated with the non-parametric log-rank test, explained in more detail in the Results section.



subsection{Bioinformatics tools}

The Cancer Cell Lines Encyclopedia 's differential expression tool was used to compare the expression levels of U-87 MG, U-251 MG, and MCF-7. The results were plotted as a heat map.

chapter{Results}

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