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Cell Culture

Types, Limitations, and Applications of Cell Suspension Culture

Cell suspension culture is a technique that allows cells to grow in liquid medium. There are several different types of cell suspension cultures. In this article, we will look at their types, limitations, and applications. This technique is often used in research and biomedical applications. It is particularly useful for studying the development of new cancer drugs.

Types, Limitations, and Applications of Cell Suspension Culture
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Table of Contents

What is Cell suspension culture?

Cell suspension culture is a technique that allows researchers to culture cells in liquid medium. In this process, cells grow in colonies that are dispersed and move within the culture flask. Sub-culturing and incubation follow until colonies reach optimal size.

Suspension cultures can be prepared from a variety of sources, including friable callus, embryo, or ground tissue. Suspensions can be manipulated by centrifugation to create fine and homogeneous suspensions, or allowed to settle and become homogeneous.

Read More: What is a Cell Culture? Application of Cell Culture?

Importance of Cell suspension culture?

Cell suspension culture is an excellent tool for examining the secondary metabolites in a variety of plants. The application of this technique has led to considerable industrial activity in the search for such molecules. In addition, cell suspension culture is used to make mutant cell clones, which can then be used in plant breeding. The clones can then be subjected to a variety of chemicals, resulting in plants with various mutations.

Types of Cell Suspension Culture

Cell suspension culture is a method used to maintain cells in culture flasks without tissue culture treatment. This method is characterized by the presence of nutrients in the culture medium and the constant addition of fresh medium.

This type of culture avoids de-detoxification of the media and overcomes the disadvantages of batch culture. The chemostat method, which adds fresh medium while harvesting the same amount of culture, is another method of maintaining steady cell growth. In this culture, nutrients such as glucose, nitrogen and phosphorous are adjusted to meet the growth limiting factors of the cells.

Read More: What is Primary and Secondary Cell Culture? Complete Guide

Cell suspension cultures are often characterized by their cell aggregate size. Depending on the size of the cells, they may produce different types of biomass. Cells that have a size of 250 um or smaller can produce higher biomass than cells with a size of 500 um. To determine the optimum size, cell suspension cultures were filtered through a stainless steel mesh of 750 um.

Suspension cultures are ideal for the production of monoclonal antibodies and cell therapy products. The resulting cells can be used in high volumes for a variety of applications. For example, CHO cells are commonly used to produce monoclonal antibodies. Researchers modified CHO cells to grow well in a suspension environment. Gene therapy products may also be produced in this way.

Applications of Cell suspension culture

Cell suspension culture is a technique that maintains cells in culture flasks without the use of tissue culture. It requires the maintenance of a constant NM of 0.2-0.5 mL/cm2 in a medium that is agitated, typically by using a magnetic stirrer or a spinning flask.

Production of secondary metabolites or recombinant proteins

It can be used for various purposes, including production of secondary metabolites or recombinant proteins. These substances are not required by plants for normal growth, but are produced during the cell metabolism process. Cell suspensions are ideal for studies in plant growth. In addition, plant cell suspension cultures are a useful tool in biotechnology. For example, plant cell suspension cultures can be used to produce plant proteins. They are particularly useful for developing drugs and diagnostic tests.

Read More: Complete guide of Genetic Toxicology

Tissue studies

Another application of cell suspension culture is for tissue studies. The resulting tissue samples are ideal for germplasm exchange. Tissue culture is also useful for studying cell differentiation and responses to pathogens. These cultures may be grown in either solid or liquid medium. It is important to use a medium with low levels of auxin and high concentrations of cytokinin.

To achieve optimum results, cell suspension culture requires careful handling. The materials and working space must be cleaned and sterilized. Cell cultures should be autoclaved after use. In addition, cross-contamination between different cell types is a major issue. Different cell types should never be processed at the same time and working areas must be cleaned between different types of cell preparations.

Limitations

  • Cell suspension culture is a method used to cultivate a wide range of cell types. In contrast to adherent cultures, which require a fixed anchorage to grow, suspension cultures are scalable. However, suspension cultures do have several limitations. First, they are not as easily passaged as adherent cultures. Second, suspension cultures do not allow for direct visualization of cells. In order to minimize these limitations, cell suspension cultures must be shaken constantly to maintain adequate gas exchange.
  • The benefits of this culture method outweigh the drawbacks. This simple culture system allows for the rapid generation of tumor cell lines and is compatible with high-throughput drug screening. However, the major limitations of this approach include low success rates, particularly for long-term passages. Moreover, there are no reliable cell viability assays available for MCTS.
  • The second limitation is that cell suspension cultures require the use of a specialized medium. The medium is usually derived from plant tissue or a variety of animal tissues. For example, the medium in which suspension cells grow should not be diluted more than 0.2 to 0.5 mL per cm2. A mechanical stirrer or a rotating spinner flask is a simple way to agitate a cell suspension culture.
  • Plant cell suspension cultures are widely used for the production of recombinant proteins and secondary metabolites. Using plant cells for this purpose has many advantages over microbes. For example, plant cells are not prone to endotoxins and carry out higher eukaryotic posttranslational modifications. In addition, plant cell suspension cultures are more cost-effective than bacterial expression systems.

New Approach in Cell Suspension Culture

Collagen is an essential component of the extracellular matrix (ECM). Its structure is composed of three a-chains that provide tensile strength to the ECM. Glycine and proline are important components of collagen that contribute to the stability of the triple helix. These triple helices aggregate into fibrils and polymerize to form fibers.

Read More: What does Adherent Cell Culture Mean? Comprehensive guide

Extracellular matrix mimic

Cell suspension cultures in a controlled extracellular matrix mimic the complex three-dimensional structure of the body’s tissues and are beneficial for high-throughput screening. The submillimeter-sized compartments have a thin alginate hydrogel shell and a collagen matrix core. The three-dimensional environment helps the cells form self-organized multicellular assemblies and is useful in high-throughput screening.

Collagen hydrogels

Collagen hydrogels are biomimetic, biocompatible, and cytocompatible. They mimic the native environment of the cell and present a native viscoelastic environment to the cells. As a result, collagen hydrogels are an excellent model for cellular microenvironments. However, there are several challenges with collagen as a protein, including its low stiffness and long-term stability. The gelation temperature and duration of time are critical in determining the mechanical properties of collagen hydrogels.

Read More: Cell Culture Media Components and Preparation- A Complete Guide

Collagen component

This new method allows researchers to measure the collagen component in cell suspension cultures with minimal effort. The suspension media is harvested directly from 96-well plates and counted using a Wallac 1205 flat-bed scintillation counter. It was validated by using the total collagen assay. The new method is a rapid and simple one-step quantitative approach to measure collagen synthesis without the use of extensive dialysis or precipitation of proteins.

Cell Suspension Culture Protocol

  • Cell suspension culture has several advantages over other types of cell culture. First, it allows for the easy passage of cells without enzymatic or mechanical dissociation. Then, it allows for high-volume cell production. Finally, the method of cell suspension culture is suitable for industrial-scale production. Since the cells are suspended in the medium, it allows for a high degree of control over the growth rate. Also, it is easy to scale up the production.
  • After this process, the cells are collected into tubes. These tubes are then centrifuged to pellet the cells. The pellets can then be stained with trypan blue to count living cells. Next, the cells are re-suspended in fresh medium. Finally, the concentration of cells can be determined using various methods of cell dilution.
  • Cell suspension culture is often used for research on cell growth, as it can be used to produce large quantities of bioactive metabolites. In most cases, cell suspension cultures require two steps, the first step is to grow the cells until they are mature, and the second step involves the stimulation of metabolite production. In the second step, reagents or environmental conditions are added to the medium to cause the cells to produce more metabolites.
  • Cell suspension culture can be used to maintain a constant growth phase, while avoiding the negative aspects of batch cultures. In addition, fresh medium is continuously added to the culture, preventing media detoxification. This overcomes the main disadvantage of batch cultures. It is also possible to use a chemostat method to control the growth rate of cells in a fixed medium.

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