Cell Culture: Cell Culture Basics

Summary

This video script offers a comprehensive guide to the fundamentals of cell culture, emphasizing the importance of quality cells, reagents, sterile techniques, and proper laboratory equipment. It outlines the essential tools found in a cell culture lab, such as a cell culture hood, incubator, and microscope, and provides tips for maintaining a sterile environment and optimal cell growth conditions. The script also covers the proper handling and storage of media and reagents, and the importance of daily cell monitoring for health and contamination. Additionally, it advises on cell transportation and authentication to ensure accurate cell culture practices.

Takeaways

• 🔬 **Quality Matters**: Successful cell culture relies on high-quality cells, reagents, and equipment.
• 👩‍🔬 **Sterile Technique**: Maintaining a sterile environment is crucial to prevent contamination.
• 🧪 **Equipment Essentials**: Basic lab equipment includes a cell culture hood, incubator, centrifuge, and microscope.
• 🛠️ **Proper Setup**: Having all necessary materials close at hand is important for efficient cell culture work.
• 🌡️ **Incubator Conditions**: Mammalian cells typically require 37°C, 5-7% CO2, and high humidity.
• 📏 **Space Management**: Keep cell culture flasks evenly spaced in the incubator to ensure proper gas exchange.
• 💊 **Media Storage**: Store media and reagents as per label instructions to prevent degradation.
• 🕒 **Reagent Warming**: Warming reagents to 37°C before use can take 10-20 minutes, depending on bottle size.
• 🧴 **Cleaning Protocol**: Clean the cell culture hood and all items within it with 70% ethanol to maintain sterility.
• 🔎 **Daily Monitoring**: Regularly check cell cultures under a microscope for health, growth, and contamination.

Q & A

What are the key factors for successful cell culture?

Successful cell culture depends on the quality of cells and reagents, sterile technique, experience, and appropriate laboratory equipment.

What is the purpose of a cell culture hood?

A cell culture hood provides a sterile environment to work with cells, minimizing the risk of contamination.

Why is it important to keep cell culture dishes and reagent bottles in a sterile environment?

Opening cell culture dishes and reagent bottles in a sterile environment prevents contamination, which is crucial for cell health and growth.

What is the typical temperature and CO2 level for culturing mammalian cells?

Mammalian cells are typically cultured at 37 degrees Celsius in the presence of 5 to 7 percent carbon dioxide with high humidity.

How should cell culture flasks be arranged inside an incubator?

Cell culture flasks should be evenly spaced inside the incubator to ensure free air movement, maintaining temperature, humidity, and proper gas exchange.

Why is it important to store media and reagents according to label instructions?

Some components degrade when left at room temperature or exposed to light, so proper storage is essential to maintain their effectiveness.

What is the recommended procedure for warming reagents before use?

Reagents can be warmed to 37 degrees Celsius, which should take 10 to 20 minutes depending on the bottle size, or used at room temperature or directly from 4 degrees Celsius storage.

What are some tips for using a water bath to warm reagents?

Bottles should be kept upright with necks out of the water. Special bottle weights or metallic bath beads can be used to keep bottles submerged without water contact.

How should items within the cell culture hood be arranged for optimal workflow?

Items should be arranged with the pipette aid in front, reagents in back, tube rack in the rear left, and a small container for liquid waste in the back right, with cell plates in the center.

Why is it important to clean the cell culture hood and items within it with 70% ethanol?

Cleaning with 70% ethanol helps maintain a sterile environment within the hood, preventing contamination and ensuring cell culture integrity.

What should be done daily to monitor the health and growth of cells in culture?

Every culture should be viewed under a microscope daily to check cell health, growth, and for signs of contamination.