The effects of the cannibinoid drug on ACEA and Hypoxia on DIPG cells

My lab report is on the effects of cannabinoid agonist ACEA on the cell cycle of DIPG cells and how Hypoxia also affects DIPG cells. I have results consisting of light microscopy cell images. Cell count and viability and two-step cell cycle assay graphs (which is pretty much the same as flow cytometry.

Images of DIPG and astrocyte cells have been taken at 24 and 48 hours. These cells have been treated with a cannabinoid agonist ACEA. The concentrations administered to the cells are 0um (negative control), 1um, 100um and positive vector control of ethanol. This was repeated 3 times for each concentration. I also have images of just DIPG cells taken at 24 hours that are in hypoxic and normoxic conditions which have also been treated with the same concentrations and controls. NOTE: The astrocytes haven’t been tested at a concentration of 1um for the 24and48images.

I would like comparisons to be made between the images:

• The controls and the concentrations. Talking about the effects on cellular proliferation and if the cells look alive or dead.
• Comparison between 24 hour and 48 hours
• Comparison between Hypoxic and normoxic cells
• The effects on DIPG cells and astrocytes. (fyi DIPG cells are cancerous and astrocytes are normal healthy brain cells but if you dont know that then I dont think this is for you.)

I have cell count and viability assays. These have been treated in the same way as the images. They have only been treated in normoxic conditions.
I used two different methods for the cell count. One uses trypan blue and one uses nuclei counter. Nucleocounter is a more accurate measuring technique and would be the optimal one to make graphs for and might be worth briefly discussing.

• I would like professional looking graphs of the cell count and statistical analysis of the significance of cell count and cell viability between the controls and 100um concentrations. I’d like to see if there’s a difference in the effects of the drug on astrocytes compared to DIPG or if both have an equally decreased cell viability at the 100um concentration.

Lastly, I have graphs which are like flow cytometry graphs called two step cell cycle assay graphs which show the percentages of cells in certain phases of the cell cycle. These have also been treated in normoxic conditions at concentrations of 0um, 1um, and 100um.

There doesn’t look like there is any difference between the graphs. So you could just write about the percentages and show there’s no significant difference in the value of the concentration.

Try and relate the discussion mainly to the effect of the cannabinoid agonist drug (ACEA) DIPG cells to the cell cycle of DIPG cells. Also, talk about anything else relevant in the discussion and if you have any questions I will respond very quickly.