Week 2 – An Overview of My Experiment

Feb 25, 2019

Last week, I went into the details on why liver cancer is so detrimental, as well as how my project would help find better treatment methods for Hepatocellular Carcinoma (HCC). In this week’s blog, I hope to explain what I’m testing and how I will be doing that.

The line of cells I use are called HepG2 cells, with “Hep” referring to liver. These cells were taken from a 15-year-old white male, who unfortunately died only a couple years later to the disease. As you know, normal cells have only 23 pairs of chromosomes, but HepG2 cells are known for their whopping 55 pairs. They are very easy to grow and easily adherent (meaning they stick to the culture wares) which makes them a good cell line to be tested on. These cells, mainly because of their cancerous characteristics, grow extremely fast. They end up using up all the nutrients in their media within 2 days, and will grow to a number from 200k to 20 million cells within that time.

Each trial in my project will have two stages: The first stage being treatment and FACS analysis, and the second stage being treatment and Western Blot. The treatment procedures will be the same, making the main difference the culture ware and number of cells. In the first stage, I use 2 12-well plates, each well holding 1 mL of HepG2 cells. With 24 wells in total, there are 24 conditions. As a small reminder from my last blog, I am testing the combination of Doxorubicin and Paclitaxel. Click on this link –> Experiment Overivew to see a diagram of all the conditions. At least 12 hours after treatment, I collect the cells, arrest their growth, and then insert them into a machine for FACS analysis. FACS analysis basically gives me 2 sets of data: The number of dead vs live cells, and cell cycle data. I will elaborate more on the data next week.

In the second stage, I take 10 conditions based on the FACS analysis results. I have highlighted those 10 conditions in the Experiment Overivew sheet. From there, I use 10 petri dishes, 1 per condition. Once I treat them the same way as before, after at least 12 hours, I collect the cells again. However, this time I must subject those cells to protein extraction and Western Blot, which basically shows which proteins are expressed.

Finally, based on both the FACS analysis data and the Western Blot data, I can determine the best Dox and Pac combination for HCC treatment.

One Reply to “Week 2 – An Overview of My Experiment”

  1. Sahithi P. says:

    Thank you for clearly and thoroughly explaining your procedures, but I think some pictures or diagrams would really help me understand it.

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