The cell count per mL in our cell culture dish was 610,000. The total cell numbe

Question

The cell count per mL in our cell culture dish was 610,000. The total cell number obtained in the 2.5 mL cell suspension we prepared from our original dish was 163.9 mL. We got 0 % of viable cells in the suspension.

Compared to the results obtained by the other groups in class. Is there any difference between those cells treated with H2O2 and those that were not?

According to the standard deviation

6 Water Treatments for Groups: 2, 3, 4, 6, 7, 10 = HAD MORE DEAD CELLS

6 Hydrogen Peroxide Treatments for Groups: 1, 5, 8, 9, 11, 12 = HAD LESS DEAD CELLS

1) Summarize your interpretation of the findings/ results from given information?

2) Include a statement about the future scientific directions suggested by your findings from the experiment. suggest future experiments using as base the results obtained in the lab?

AND IF POSSIBLE CAN YOU HELP WITH A GRAPH FOR % MORTALITY VS. GROUP NUMBERS

1 2 3 4 5 6 7 8 9 10 11 Number of cells 165,000 1,285,000 610,000 4,500,000 505,000 1,270,000 1,650,000 10,800,000 15,187,500 570,000 1,525,000 895,000 ml for 100,000 cells 606 ml 77.82 ml 163.9 ml 22 ml 198.02 ml 78.74 ml 60.60 ml 231.48 ml 125 ml 175.4 ml 98.36 ml 114.94ml % mortality 12.12% 0.78 % 0% 1.00% 8.9% 3.15% 6.97% 37.03% 2.88% 7.02% 0.079% 1.70%

Explanation / Answer

1)   Summarize your interpretation of the findings/ results from given information?

One possible explanation of what you are seeing in your experiments could be that hydrogen peroxide treatments depending on the concentration of hydrogen peroxide can cause death to cells or not. For this reason, are many scientists looking and trying to determine lethal doses of hydrogen peroxide in cells. From what you observe in the given experiments where the groups of cells treated with hydrogen peroxide had less dead could depend in the hydrogen peroxide doses, the exposure time of incubation with hydrogen peroxide which could vary depending on the cell type. An average found where cytotoxicity takes place is between 4 – 24 hours. So that could be a possible reason why are you seeing that hydrogen peroxide treatment is not showing the dead cells expected, probably you need more time to see the results of the cytotoxicity.

On the other hand, cells treated with water showed more dead cells probably because the water was not on the proper concentration of solutes to establish an isotonic solution, causing a cell swelling like hypertrophy problem and then causing the apoptosis of the cells. Cells have the ability be impermeable to ions and other solutes, but like all phospholipid bilayers, they are somewhat permeable to water. Water tends to move across a membrane from a solution of low solute concentration to one of high. Cell swelling is an early change that occurs in most types of acute injury and which can be a prelude to more drastic changes. Cell swelling results when the cell loses its ability to precisely control the movement of ions and water into and out of the cytosol. Causing hypertrophy which is the increase in the cell size and results from an increase in the amount of cytoplasmic organelles.

2)   Include a statement about the future scientific directions suggested by your findings from the experiment. suggest future experiments using as base the results obtained in the lab?

For future work, everything will depend on what are you looking for, what are your objectives. For example, if your main objective is to determine a lethal dose of hydrogen peroxide compared to water solution, probably you will need to do experiments with higher concentrations of hydrogen peroxide compared to the concentration used in the actual experimentation. So, you can observe a higher cells death compared to cells treated with water solution. Time is also important; the incubation time expose with hydrogen peroxide plays an important role is not the same to compare your results one hour after you dilute your cells with hydrogen peroxide solution to four hours later where the ability of hydrogen peroxide to cause death to cells is more visible.

If that is not the main objective to find the cytotoxicity of the cells using hydrogen peroxide, probably that hydrogen peroxide concentration is adequate, but you need to change your solute concentrations in water in order to keep an isotonic solution where cells are not stressed by other factors and avoid cell swelling. So, you can compare which treatment is the more adequate to keep the cells viable.

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