Overview

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• Source: Microbioz India


• Date: 03 Sep,2023

Growing and maintaining cells in a laboratory-controlled setting is known as cell culture. Numerous scientific disciplines, including cell biology, cancer research, drug discovery, and tissue engineering, greatly benefit from this method.

Cell Culture Techniques:

Cell Line Selection:

Researchers choose a specific cell line based on their research objectives. Cell lines can be primary (isolated directly from tissues) or immortalised (can divide indefinitely).

Sterile Technique:

Maintaining sterility is critical to avoid contamination. Researchers use a laminar flow hood or biosafety cabinet to work with cells under sterile conditions. Proper handwashing, lab coats, gloves, and disinfectants are essential.

Cell Culture Media:

Cells are grown in specialised culture media that provide essential nutrients, growth factors, and supplements. The choice of media depends on the cell type and research objectives.

Also read:

Pluripotent Stem Cell Culture Media: All you need to know

Subculturing:

Cells need to be regularly passed to prevent over confluence, which can lead to cell death. Subculturing involves detaching cells from the culture vessel and seeding them into new flasks or plates.

Cryopreservation:

Low temperatures (-80°C or in liquid nitrogen) allow for the long-term storage of cells. Cells are protected from harm caused by freezing and thawing by being treated with cryoprotectants.

Cell Counting:

Various methods are used to determine cell concentration, including hemocytometers, automated cell counters, and flow cytometry.

Quality Control:

Regularly check cells for morphology, growth rate, and contamination. Mycoplasma testing is essential to detect bacterial contamination.

Common Errors in Cell Culture:

Contamination:

Contamination with bacteria, fungi, or other cell lines can lead to unreliable results. Maintaining sterile technique and routine testing for contaminants is crucial.

Overgrowth and Senescence:

Allowing cells to become too confident or passing them too many times can lead to senescence or cell death.

Media and Reagent Quality:

Using expired or low-quality media, sera, or reagents can impact cell health.

Incorrect Temperature and CO2 Levels:

Cells often require specific temperature and CO2 levels. Failure to maintain these conditions can affect cell growth.

Inadequate Documentation:

Keeping detailed records of passages, media formulations, and experimental conditions is essential for reproducibility.

Equipment Used in Cell Culture:

Cell Culture Incubator:

Maintains temperature, humidity, and CO2 levels for cell growth.

Laminar Flow Hood/Biosafety Cabinet:

Provides a sterile work environment for handling cells.

Microscope:

Used for observing cell morphology and confluence.

Pipettes and Pipette Tips:

For accurate measurement and transfer of liquids.

Cell Culture Dishes, Plates, and Flasks:

Containers for cell growth and experiments.

Centrifuge:

Used to pellet cells or separate cellular components.

Cryopreservation Equipment:

Includes cryovials, liquid nitrogen tanks, and controlled-rate freezers for cell storage.

Cell Counting Equipment:

Hemocytometers, automated cell counters, and flow cytometers.

Autoclave:

Sterilizes equipment and media.

Microbiological Safety Cabinet:

Used for working with hazardous materials like certain viral vectors.

Water Bath:

Used to warm media and reagents.

Cell Scrapers and Dissociation Reagents:

Tools for detaching cells from culture vessels.

FAQS ON Cell culture:

 What are cell culture techniques?

Cell culture is the maintenance and growth of cells of many organisms in vitro in special containers created under temperature, humidity, nutrients, and disease-free conditions. In general terms, cells, tissues and organs that are isolated and stored in the laboratory are considered tissue products.

 Cell culture in microbiology

Cloning and generation of recombinant proteins are two common molecular biology applications of microbial cell culture. In the medical field, they play an important role in the detection, diagnosis, and naming of potential pathogens. Using microbial culture, cells can be grown and divided in a lab under precise laboratory conditions.

Applications of Cell Culture

Cell culture is one of the most important tools in cell and molecular biology; It provides excellent models for studying the physiology and biochemistry of cells, the effects of drugs and toxins in cells, mutation and carcinogenesis.

Types of cell culture

For the growth of cells in culture, there are two methods of cell culture, such as a single layer of cells on a substrate (i.e. culture) or free-floating in the medium (the traditional route).