Cell Culture techniques - Histology

What is Cell Culture?

Cell culture refers to the process by which cells are grown under controlled conditions, typically outside their natural environment. This technique is crucial in histology for studying the structure and function of cells, tissues, and organs.

Types of Cell Culture

There are several types of cell culture techniques, which can be broadly categorized into primary culture and secondary culture. Primary culture involves the direct isolation of cells from tissues, while secondary culture involves the subculturing of cells from a primary culture.

What are the Key Components of Cell Culture Media?

The growth medium is essential for cell survival and proliferation. It typically contains amino acids, vitamins, minerals, glucose, and serum. The medium also needs to maintain the appropriate pH and osmolarity.

Why is Sterility Important?

Maintaining sterility is crucial to prevent microbial contamination. Contaminants can outcompete the cells and lead to erroneous results. Techniques such as working in a laminar flow hood and using antibiotics can help maintain sterility.

How are Cells Isolated for Culture?

Cells can be isolated using enzymatic digestion methods like trypsinization or mechanical methods like tissue dissociation. The choice of method depends on the tissue type and the desired cell yield.

What are the Commonly Used Cell Lines?

Commonly used cell lines include HeLa cells, CHO cells, and 293T cells. These cell lines are well-characterized and widely available, making them ideal for various experimental setups.

How are Cells Subcultured?

Subculturing involves transferring cells from one culture vessel to another to prevent overconfluency and maintain healthy growth. This typically involves trypsinization to detach adherent cells and then reseeding them into fresh medium.

What are the Applications of Cell Culture in Histology?

Cell culture techniques are used in histology for various applications, including drug testing, gene expression studies, and tissue engineering. They allow researchers to observe cellular responses in a controlled environment.

How is Cell Viability Assessed?

Cell viability is assessed using methods like trypan blue exclusion and MTT assay. These tests help determine the percentage of live cells in a culture, which is crucial for experimental accuracy.

What are the Challenges in Cell Culture?

Challenges include contamination, genetic drift, and maintaining the phenotype of primary cells. Overcoming these challenges requires strict adherence to protocols and regular monitoring of cultures.

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