Cell Culture 101 1

Summary

In this educational video, molecular researcher Laura from the University of Toronto introduces the fundamentals of cell culture, specifically focusing on the MCF7 breast cancer cell line. She explains the importance of maintaining a sterile environment using airflow hoods and ultraviolet lights to prevent contamination. Laura demonstrates how to prepare cell culture media with supplements like fetal bovine serum and antibiotics, and how to passage cells using trypsin-EDTA. She also discusses the concept of cell confluence and the process of treating cells with estradiol, highlighting the steps involved in cell culture for scientific research.

Takeaways

• 🔬 Laura is a molecular researcher at the University of Toronto introducing cell culture basics.
• 🌟 Cell culture involves growing eukaryotic cells in vitro for scientific research purposes.
• 🧫 The MCF7 breast cancer cell line is used for demonstration in this tutorial.
• 🔄 Maintaining a sterile environment is crucial in cell culture to prevent contamination.
• 🌬️ Airflow hoods and ultraviolet lights are used to maintain sterility in the cell culture area.
• 💧 Cell culture media, like DMEM, provides essential nutrients for cell survival.
• 💉 Fetal bovine serum is added to the media to supply growth factors for cell growth.
• 🛡️ Antibiotics like penicillin and streptomycin are used to prevent bacterial contamination.
• 🧑‍🔬 The process includes steps like rinsing cells with PBS and using trypsin to detach them from the culture vessel.
• 📊 Cell confluence is an important measure, indicating how densely cells cover the culture plate.
• 🔄 Cells are passaged before reaching 100% confluence to avoid contact inhibition and cell death.
• 🧪 Post-passaging, cells are treated with substances like estradiol for experimental purposes.

Q & A

What is cell culture and why is it important in scientific research?

Cell culture is the growth of eukaryotic cells in vitro for scientific purposes. It is important because it allows researchers to study cell behavior, growth, and response to various treatments in a controlled environment.

Why is maintaining a sterile environment crucial in cell culture?

A sterile environment is crucial to prevent contamination from mold or bacteria, which could invalidate the results and require researchers to start the experiment again.

How do airflow hoods help maintain a sterile working environment in cell culture?

Airflow hoods maintain a sterile environment by generating a current of air that is pulled down into the hood and back through a system of filters, ensuring the air inside the hood is sterile.

What is the role of ultraviolet lights in a cell culture hood?

Ultraviolet lights mounted on the back of the hood irradiate the working surface, further reducing the potential for contamination.

What is the primary function of cell culture media like DMEM?

Cell culture media, such as DMEM, provides a balanced salt solution, pH balance, and essential nutrients necessary for cell survival.

Why is fetal bovine serum added to the cell culture media?

Fetal bovine serum is added because it contains growth factors that stimulate the growth of the breast cancer cell lines being cultured.

What is the purpose of supplementing the media with penicillin and streptomycin?

The supplementation with penicillin and streptomycin acts as a prophylactic measure to prevent any potential bacterial contamination in the cell culture.

How is the cell culture bottle prepared before use in the hood?

The bottle is prepared by spraying it with a 75% ethanol solution to disinfect the surface before placing it in the cell culture hood.

What is the function of trypsin EDTA in cell culture?

Trypsin EDTA is a digestive enzyme that digests external cellular proteins, allowing cells to lift off from the bottom of the culture vessel and be suspended in the media for passaging.

Why are cells passaged before they reach 100% confluence?

Cells are passaged before reaching 100% confluence to prevent contact inhibition, a phenomenon where cells stop dividing and may begin to die when they are too closely packed together.

How does the cell culture incubator contribute to the growth of cells?

The cell culture incubator maintains optimal conditions of 37 degrees Celsius and 5% CO2, simulating the internal environment of the human body, which is conducive to cell growth.