Name: growing cells in the lab
Uploaded: 2023-04-30T11:57:20.000Z
Duration: 1 min 21 s
Channel: JoVE

Cell culturing is a technique of growing eukaryotic cells in a laboratory setting.
It provides a model system to study the physiology of cells outside their natural environment.
When cells from a tissue sample are grown directly in culture medium, the resultant cell cultures are called primary cultures.
Such cells need artificially controlled conditions, to help them grow, such as correct cultureware, depending on if they are adherent or suspension cultures; an optimal temperature; a suitable growth medium that can provide essential nutrients, like amino acids, vitamins, salts, and growth factors; and a pH buffering system.
As the primary cell adherent culture multiplies, it gradually occupies the entire surface area of the culture dish.
At this stage, the cells must be treated with a solution of protease and chelating agent, such as Trypsin-EDTA, to loosen the cell-substrate and cell-cell interactions to obtain single cells.
A small number of these cells can then be subcultured into a new dish containing a fresh growth medium to facilitate continued growth. Such subcultured cells form secondary cultures.

cell culture

Cell Culture

Most vertebrate cells grow in vitro attached to a substrate as a monolayer, called adherent cultures. The flasks and plates used to grow cells are chemically treated to facilitate cell attachment. However, a few cell types, such as hematopoietic cells, can grow in a suspension. In contrast to adherent cultures, suspension cultures can grow in non-treated cultureware using magnetic stirrers or spinner flasks to agitate the culture media

Culture conditions
The growth medium is a crucial component for the optimal growth of cells. Previously, natural growth media like blood plasma, chicken embryo extracts, and amniotic fluid were used. In recent times, however, synthetic media such as Dulbecco&#39;s Modified Eagle Medium (DMEM) and Roswell Park Memorial Institute (RPMI) are preferred. These basal media contain amino acids, vitamins, inorganic salts, and a carbon source such as glucose. They are supplemented with fetal bovine serum (FBS) as a source of growth and adhesion factors, hormones, lipids, and minerals. Antibiotics are added to the growth media to prevent microbial contamination, and culturing protocols are performed under aseptic conditions.
The pH of the medium is usually maintained at 7.4 for mammalian cells. The growth medium also contains a buffering system such as sodium bicarbonate with exogenous CO2 for pH regulation. A pH indicator like phenol red is incorporated into the culture medium to indicate pH changes. An optimum temperature of 37&#8451; and a CO2 concentration of 5% are maintained in the incubator.

Plant cell culture
Like animal cells, plant cells can also be cultured in vitro. Plant tissue, known as explant, is cultured in a nutrient medium containing plant growth hormones, micronutrients, and a carbon source. Relative proportions of the plant growth hormones auxins and cytokinins in the medium decide the tissue type developed in the culture. A balanced ratio of these hormones results in a mass of undifferentiated cells called the callus. By altering the hormone ratio, the callus can be differentiated into root or shoot, thus generating a complete plant. Explants are usually cultured on a solid nutrient medium, while single-cell suspensions can be grown in liquid media. Growth conditions like temperature, light intensity, and photoperiod govern the plant growth in culture. Plant cell cultures are used to produce improved hybrid plants and conserve endangered plants.&#160; Explant cultures are also used in the large-scale production of plant-derived products.

Most vertebrate cells grow in vitro attached to a substrate as a monolayer, called adherent cultures. The flasks and plates used to grow cells are ...

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Analyzing Cells and Proteins

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growing cells in the lab

Growing cells in the lab depends on the right culture type and the right growth conditions. Most vertebrate cells grow attached to a surface as adherent cultures. These cells are grown in flasks and plates that are chemically treated to help the cells attach. Some cells, such as hematopoietic cells, grow in suspension instead. Suspension cultures can be kept in non-treated cultureware with magnetic stirrers or spinner flasks to keep the medium moving.

The growth medium is a key part of successful cell culture. Earlier, natural media such as blood plasma, chicken embryo extracts, and amniotic fluid were used. Today, synthetic media such as Dulbecco's Modified Eagle Medium (DMEM) and Roswell Park Memorial Institute (RPMI) are preferred. These basal media contain amino acids, vitamins, inorganic salts, and a carbon source such as glucose. They are also supplemented with fetal bovine serum (FBS), which provides growth and adhesion factors, hormones, lipids, and minerals. Antibiotics help prevent microbial contamination, and the work is carried out under aseptic conditions.

Cell cultures also need tightly controlled physical conditions. For mammalian cells, the medium is usually kept at pH 7.4. A buffering system such as sodium bicarbonate works with added CO2 to regulate pH. Phenol red is included as a pH indicator so changes in acidity can be seen. An incubator typically maintains 37c and 5% CO2.

Plant cell culture follows a similar idea but uses plant tissue called an explant. The explant is placed in a nutrient medium that contains plant growth hormones, micronutrients, and a carbon source. The balance between auxins and cytokinins determines what tissue develops. A balanced ratio produces a mass of undifferentiated cells called callus. Changing the hormone ratio can turn callus into roots or shoots and eventually into a complete plant.

Plant explants are usually grown on a solid nutrient medium, while single-cell suspensions can grow in liquid media. Growth also depends on temperature, light intensity, and photoperiod, which is the light-dark cycle. Plant cell cultures are useful for making improved hybrid plants, conserving endangered plants, and producing plant-derived products on a large scale.

Growing Cells in the Lab

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Growing cells in the lab depends on the right culture type and the right growth conditions. Most vertebrate cells grow attached to a surface as adherent cultures. These cells are grown in flasks and plates that are chemically treated to help the cells attach. Some cells, such as hematopoietic cells, grow in suspension instead. Suspension cultures can be kept in non-treated cultureware with magnetic stirrers or spinner flasks to keep the medium moving. The growth medium is a key part of...

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key terms and concepts

Learn how animal and plant cells are grown in culture, from adherent and suspension cells to media, pH control, and plant callus formation.

Growing Cells in the Lab | JoVE K12

cell biology

JoVE Core Cell Biology illustrates complex cellular processes in easy-to-understand yet comprehensive animated video lessons. This collection focuses on the essential components and functions of cells, including organelles and membranes, cellular transport, and cell energetics. It is complemented by scientist-in-action videos that demonstrate the application of cell biology in real-world research experiments.

Growing Cells in the Lab

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