Treating U87 EGFP Glibolastoma Cells with Nickel Sulfate!

Part of the reward of being a Biomedical Engineer is the opportunity to explore fascinating and cutting-edge work that teaches me new skills and exposes me to unique challenges every semester. Throughout my journey, I’ve learned coding, worked with circuits, conducted detailed statistical analyses, and performed rigorous statistical tests. However, this semester, I had the chance to do something I had never experienced before. I worked with cancer cells. Yes, you read that right! I worked with cancer cells in a wet lab environment!

What is a wet lab, you ask? It’s a controlled environment where you work with potentially hazardous cell cultures and substances while following strict safety protocols, such as wearing gloves, goggles, lab coats, and disinfecting surfaces with ethanol to avoid contamination. Mistakes can introduce bacteria or other pathogens that could destroy your cultures, making careful adherence to procedure critical. Despite the challenges, working in a wet lab is an incredibly rewarding experience that not everyone gets to have. To share this unique experience, I want to provide some insight into what a typical experiment looks like in a wet lab, so those who haven’t had the opportunity can get a feel for it. Trust me, you’ll find it fascinating!

One of the experiments I conducted this semester involved treating Glioblastoma (U87-EGFP) cells with varying concentrations of Nickel Sulfate (NiSO₄) to assess its effects on cell viability. Intuitively, I hypothesized that increasing concentrations of Nickel Sulfate would be progressively toxic to these cells. Sure enough, that’s exactly what happened. The coolest part? I didn’t even need to run complex calculations to see the effects! By observing the cells under a microscope, I could identify the toxicity based on changes in cell morphology and network structure. Let me explain how this works.

When cancer cells are healthy, they exhibit a spherical shape and form densely interconnected networks, similar to what we see in normal, healthy cells. However, as cells are damaged, these networks weaken, and their shapes become less defined and less spherical. These morphological changes occur because interactions between the cells’ mitochondria and cytoskeleton are disrupted. Observing these changes during the experiment allowed me to visually interpret the effects of Nickel Sulfate on the cells. Below, I’ve described and analyzed the experiment’s results to demonstrate how the process unfolded in real time.

Figure 1A. Appearance of U87 – EGFP cells, after addition of 0 mM of NiSO4. Cells appear to be healthy, as they are exhibiting a bright color, are elongated in shape, and are forming an interconnected network like structure.

Figure 1B. Appearance of U87 – EGFP cells, after addition of 0.0625 mM of NiSO4. Cells still appear to behealthy, but they are slightly less elongated, and are slightly less bright in color, as compared to when 0 mM ofNiSO4 was added. The cells do still appear to be interconnected, though.

Figure 1C. Appearance of U87 – EGFP cells, after addition of 0.125 mM of NiSO4. Cells appear to be less healthy, and are significantly less elongated and bright, compared to when 0 or 0.0625 mM of NiSO4 wasadded. Cells also appear to be significantly less interconnected, as the network structure is weakening. The addition of NiSO4 at this concentration seems to be having a significant impact on the structure of NiSO4, indicating that the IC50 concentration is being approached.

Figure 1D. Appearance of U87 – EGFP cells, after addition of 0.25 mM of NiSO4. Cells appear to be significantly less healthy, and there is hardly any presence of elongation, compared to when 0, 0.0625, or even0.125 mM of NiSO4 was added. There also appears to be hardly any network structure left between the cells, as all interconnections are not visible. The cells are also significantly less bright, and are extremely dim. The significant structural changes that arise from the addition of this concentration of NiSO4 indicate that the IC50 concentration has been passed.

Figure 1E. Appearance of U87 – EGFP cells, after addition of 0.50 mM of NiSO4. Cells appear to be significantly less healthy, and do not have any visible interconnected network structure. Any slight presence of anetwork structure seen when 0.25 mM of NiSO4 was added also appears to be gone. Now, there appears to be no network structure.

Figure 1F. Appearance of U87 – EGFP cells, after addition of 1 mM of NiSO4. Cells appear to be significantly less healthy, as there is no elongation present. There is also no network structure left between the cells. No interconnections appear to be visible. The cells are in a completely opposite state to how they were, from a color perspective, when they were treated with no NiSO4. Given the permanent changes in structure and morphology of the cells, it is likely that the addition of 1 mM NiSO4 has the maximum possible impact on the cells. In other words, it is likely adding any more NiSO4 will have a marginal impact, if any.

Overall, observing these morphological changes was an eye-opening experience for me as it taught me to analyze cellular changes, in a relatively simple way, without relying on quantiative measurements or complicated formulas, something that I never knew that you could do as an Engineer! It also taught me that cancer is not as invincible as it is made out to be and it can indeed be quashed. This is something that I never knew until I did this experiment, and it gave me a lot of hope for the future of cancer treatment!

Reading this post, I hope you got inspiration to work in a Wet lab too one day as you see all the cool stuff that you can do/accomplish. Sometimes, you do not have to be a fancy scientist at the NIH or the CDC to make a difference. Occasionally, just the Georgia Tech student can accomplish some pretty cool stuff!

 

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