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Cell culture is a general term that describes the process of growing cells in an artificially created environment. For life science laboratories, maintaining mammalian cells is a regular task in the cell analysis workflow. It is a consistent method in molecular and cell biology to create desirable cells for analysis, with applications in cell-based assays, as a model system for diseases and for drug screening and development.

Mammalian cells such as Animal and human cells can be grown outside the organism given the right physical conditions, medium, and growth factors. Cells may be obtained from live tissue directly to create primary cell cultures, or derived from an existing cell line. A primary cell culture is either obtained or created through enzymatic or mechanical methods, and grown to full cell confluence (fully covered) before a cell line can be established. At this stage, the primary cell can be subcultured in a process known as passaging. The culture at full cell confluence is transferred to new media, enabling further propagation and the creation of a large number of cells of the same line or strain. These subcultures can be further used experimentally as samples to test how cells can be manipulated biologically, chemically or physically.

Types of Cell Cultures

Adherent cells

Also known as anchorage-dependent or monolayer cells, are grown in cell culture medium while attached to the bottom of the flask. Adherent cells have the added necessity of a removal step for processes such as passaging, where cells must be physically or chemically detached from the flask. Most tissue cells are adherent.

Adherent cells have the added necessity of a removal step for processes such as passaging, where cells must be physically or chemically detached from the flask.

Suspension cells

Suspension cells are free-floating within medium, and can be used for bulk applications due to its ability to use up all available media quickly. However, suspension cells need to be agitated by a shaker.

 

 

Suspension cells are free-floating within medium, and can be used for bulk applications due to its ability to use up all available media quickly.

Applications

  • Basic biology and biochemistry
  • Model system for drug discovery and development
  • Study the interaction between cells and disease causing agents in pathology
  • Cancer research
  • Stem cell research
  • Virology
  • Cytotoxicity testing
  • Production of vaccines, monoclonal antibodies, cell and gene therapy products

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Cell Incubation

Mammalian cells can be cultured inside climate-controlled chambers known as CO2 Incubators. The goal of these incubators is to artificially mimic the in-vivo physical environments favorable to cell growth while minimizing microbiological and physical risks to damage of the cells growing inside.

A CO2 incubator controls the following physical parameters:

  • Temperature
  • Osmotic Pressure
  • pH level (through CO2 levels)
  • Optional: O2 levels for hypoxia condition studies

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Media and Serum

Cell culture is a basic laboratory technique that allows the maintenance and growth of living cells outside of the animal or plant where they occur naturally. Successful cell culture provides cell population-optimized base media, supplements, and growth factors that mimic in vivo conditions.

The success of cell culture depends on the optimization of the culture conditions with appropriate levels of:

  • Gases (O2, CO2), pH, pressure, and temperature
  • Suitable media to provide the needed nutrients, minerals, salts, and amino acids
  • Effective growth factors to maintain phenotype and overall cell health
  • Proper cell attachment substrates for adherent cell cultures

 

Special Conditions Cell Cultures

Live cell imaging is the time-lapse microscopy of dynamic processes in living cells. It enables observation of cell-cell interactions, the behavior of single cells, and the dynamics of cell organelles or cellular molecules. Several imaging techniques can be applied, such as phase contrast microscopy, fluorescence and confocal microscopy, multiphoton microscopy, light sheet microscopy, or even TIRF and super-resolution microscopy. Live cell imaging can be performed by putting a microscope inside a CO2 incubator, or putting a CO2 incubator under a microscope, depending on preference and application.

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The majority of cells in living tissue grow in a three-dimensional microenvironment, where they communicate and interact with each other and their surroundings. Animal cells are embedded in the extracellular matrix (ECM), which is composed of proteoglycans and fibrous proteins (mainly collagen, elastin, and fibronectin). This complex, dynamic, and tissue-specific 3D structure provides physical scaffolding for the cells and initiates cues that influence cell differentiation and behavior. 

Cells behave differently inside a 3D gel matrix compared to a 2D environment. In many cases, a 3D environment reflects the in vivo situation more accurately. This should be considered when analyzing cell behavior, differentiation, response to drug treatment, and gene and protein expression.

Not surprisingly, many cell cultures approaches have been adapted to a 3D environment. This includes drug screenings that use spheroids and organoids, which are indispensable nowadays as tumor models.

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Liquids are a crucial component of every living species. Liquid flow causes shear stress, a mechanical force that influences the cell morphology and behavior in many ways. Working under flow conditions can be especially important when using cells that occur in biofluidic systems, such as endothelial or epithelial cells. Examples are:

  • vascular endothelial cells that form the inner layer of blood vessels,
  • lymphatic endothelial cells that form the inner layer of lymphatic vessels,
  • epithelial cells of the kidney and the lung.

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