Dr. & Prof. Sa-ik Bang

This internship aimed to provide cell culture practice, with a focus on HepG2 cell cultivation and seeding, the preparation of BSA-conjugated PA mixtures, the Ionpath-palmitate labeling method, and checking the Pal AC effect in liver cells. I also wanted to learn how to work as a team in the lab, improve my collaboration and communication skills, build resilience while handling experiments not yielding expected results, and be exposed to ethical considerations that matter when conducting scientific research.

1. Cell Culture and Maintenance

• I was responsible for performing cell culture of HepG2 cells. This involved handling frozen cells from liquid nitrogen, thawing them, and establishing new cultures in a controlled environment.

• I learned how to centrifuge cells, add appropriate media, and ensure cell viability in the incubator.

2. Media Preparation

• Prepared different types of media, including DPBS, trypsin, and cell culture media, ensuring that they were sterile and ready for experiments.

• During the experiments, I was required to balance the composition of the media in each step to sustain optimal conditions for cell growth.

3. Cell Seeding and Counting

• Performed cell seeding to suspend cells at a known concentration for future experiments.

• I utilized trypan blue dye and a hemocytometer to calculate the concentration and viability of cells and learned to adjust this concentration for experiments using mathematical formulas.

4. Preparation of BSA-Conjugated PA Mixture

• I prepared bovine serum albumin (BSA)-conjugated palmitic acid (PA) mixtures to study their impact on HepG2 cells.

• The process involved creating specific concentrations of PA with ethanol and BSA, ensuring that the conjugation would allow the observation of PA's effects on the cells.

• I conducted multiple rounds of mixing and filtration to finalize the BSA-PA conjugation.

5. Lab Equipment Handling and Sterile Practices

• Gained experience using essential lab equipment such as water baths, micropipettes, centrifuges, microscopes, and cell counters.

• Strict adherence to sterile techniques was required to ensure no contamination of cell cultures. This included regularly cleaning the workbench with ethanol, handling equipment properly, and managing the clean bench environment.

6. Analytical Skills

• In each experiment, I was responsible for analyzing the data, whether it was counting cells under a microscope, calculating the amount of media needed, or adjusting parameters during cell culture.

• I monitored the cell conditions closely and followed up on whether the cells survived and thrived under experimental conditions.

7. Recording and Documentation

• Detailed all observations, procedures, and outcomes in lab reports for future reference. Properly labeling Petri dishes and vials with information such as passage numbers and dates was crucial to my work, ensuring traceability and accuracy throughout the experiments.

Sterile Techniques And Cell Culture

Coming from a non-cell-based background, I learned to appreciate the value of proper execution when it comes to procedures directly related to cell culture like thawing, re-suspending, and keeping HepG2 cells in ideal conditions. By the time I finished, though (having washed my hands at least 200 times), I could use every piece of equipment in there — micropipettes, centrifuges and understand how to keep everything sterile so nothing got contaminated. The process made me more detail-oriented and emphasized the need to maintain a clean working environment in the lab.

Precise Mixing Measurements and Calculations — Dilution Ratios

Proper pipetting and calculations were the keys in the preparation of media & cell mixtures. I honed my math abilities in tasks such as cell counting and dilutions which demanded accuracy. This made me realize how important it is to be accurate from the beginning to the end of an experiment to gain trustworthy results.

Mission Challenges and Adapt to Changes

I quickly became fluent in reading between the lines. Learning to troubleshoot has taught me so much about adjusting protocols when my first experiments fail. I learned to be flexible and resilient through these challenges which made me a better problem solver and allowed me to adjust when unexpected things happened.

Analysis and Interpretation of Data

The significance of understanding and analyzing experimental data, especially cell counts and viability, cannot be underestimated. I was able to develop my analysis again and become concise in how I present my findings, written down or verbally, as a result of this experience.

Design and Plan for the Work of Experiment Based on SYNTAX Configuration

Thus, among other lessons, I also learned the importance careful experimental design when preparing BSA-conjugated palmitic acid mixture. When we think about how to plan each step and what trouble could happen, I wonder if it is possible for a successful result not to be obtained.

Sharing and Notification

I also learned teamwork and communication from other interns and lab members. Through this experience, I was taught to distribute lab duties fairly and help my team members in any way possible so that everyone could succeed, further convincing me of the critical importance of teamwork within scientific research.

Building Ethical Resilience

These things took patience and persistence throughout this internship. I spent time thinking about cancer research ethics and appreciating the necessity of conducting experiments safely. Being through this endeared me to a much broader spectrum of how ethical research should be conducted.