In today’s world, animal cell culture technology has become a vital part of the life sciences. It provides a platform for studying regulation, proliferation and differentiation, as well as genetic manipulation. To be successful, it requires certain technical skills. This chapter explains the fundamental techniques and applications of animal cell culture.
Keywords: Cell culture technology for animal research, including applications to cell culture, cell lines and viability. To be successful, it requires special technical skills. This chapter explains the fundamental techniques and applications of animal cell culture.
What you can expect to learn?
This chapter covers the fundamentals of animal cell culture and the latest applications. This chapter aims to guide students to the most important areas of cell culture. It also demonstrates an understanding of basic cell culture concepts and how to handle cell lines. This chapter provides information on cell culture methods, serum use, viability assays and the translational importance of cell cultures.
Cell culture refers to the cultivation of cells in an artificial environment. Cells can be derived from multicellular organisms, established cell lines, or established strains. Animal cell culture was a well-known laboratory technique in the 20th century. However, the idea of keeping live cells separate from their tissue source was first discovered in the 19th Century. Animal cell culture is a key tool in life sciences research in areas that offer potential economic value and commercialization. Scientists can now work with many cells in controlled environments thanks to the development of basic culture media. This has helped them advance their understanding of cell growth, differentiation, growth factors and the mechanisms that underlie normal cell functions. High cell density culture conditions and bioreactors have been investigated using new technologies.
Biotechnology products, such as viral vaccines, are dependent on the mass culturing and culture of animal cell lines. Although many simpler proteins are being produced using rDNA in bacterial cultures, more complex proteins that are glycosylated (carbohydrate-modified) currently have to be made in animal cells. Cell culture research focuses on the effects of culture conditions on viability, productivity, as well as the consistency of post-translational modification such glycosylation. These are all important factors for the biological activity recombinant protein. Recombinant DNA (rDNA), which is used in animal cell culture, produces biologicals such as anticancer agents, immunobiologicals (interleukins and lymphokines (mABs), hormones, and monoclonal antibodies (mABs).
The use of animal cell culture in research has been widespread, from basic to more advanced. It is a model system that has been used in many research projects:
1. Basic cell biology, cell cycle mechanisms and specialized cell function are all studied.
2. To test for toxicities in order to determine the effects of new drugs.
3. Gene therapy is a method of replacing nonfunctional genes by functional gene-carrying cell lines.
4. The diagnosis of cancer cells and the role of chemicals, viruses, radiation, and radiation in them.
5. Production of pharmaceutical drugs, mABs and vaccines.
6. Production of viruses for vaccine production (e.g. chicken pox and polio),
Today, mammalian cell cultivation is an essential prerequisite for the production of biological therapeutics like hormones, interferons and clotting factors.
Go to: The development of animal cell culture
The early 20th century was the time when mammalian cell culture technology began. These cultures were initially created to study cell culture development and normal physiological events, such as nerve development. Ross Harrison, in 1907, demonstrated the first in vitro nerve fiber growth. But animal cell culture was first performed on an industrial scale in the 1950s. The need to cultivate large numbers of cells was evident after the emergence of major polio epidemics in the 1940s and 1950s. A de-activated virus vaccine for polio was the first product to be developed from cell cultures. When provided with nutrients and growth factors, many animal cells can be stimulated to grow outside their original tissue or organ.
In vitro growth requires that the culture conditions do not replicate in vivo conditions in terms of temperature, pH and CO2, O2, nutrition, or osmolality. The cultured cells need sterile conditions, as well as a steady supply and complex incubation conditions. The medium is an important factor in the growth of culture medium cells. Animal cells can be cultured in either natural media or artificial media, depending on their needs. The most crucial step in animal tissue cultivation is the culture medium. It depends on the cell type that needs to be cultured in order to produce pharmaceutical products or cell growth differentiation. There are several media options that provide a variety of benefits to cell culture, including serum-containing media and serum-free media. The development of cell lines is dependent on the availability of sterile conditions.
In the laboratory, cells from many different organisms and tissues can be grown. Cell culture used to be used to study cell growth, requirements for growth, cell cycles, and cell structure. Homogenous cultures derived from primary cells are now useful tools for studying the biology and origin of cells. For the production of artificial tissues, it has been helpful to use histotypic and organotypic cultures that replicate the respective organs/tissues.
How do cell cultures are made?
Three methods are commonly used to start a culture with animals.
Organ culture
In vitro organ culture can be initiated using whole organs taken from embryos and partial organs. The cells used in organ culture retain their in vivo architecture and differentiated character. Cell proliferation is restricted to the explant’s periphery and they don’t grow quickly. These cultures can’t be propagated over long periods of time so a new explanation is needed for each experiment that results in interexperimental variation in terms reproducibility or homogeneity. Organ culture can be used to study the functional properties of cells (production and release of hormones), and to examine the effects of other agents (such drugs or other micro- and macro molecules) on organs that are anatomically separated in vivo.
Primary explant Animal Cell Culture
You can keep fragments made from animal tissue in many different ways. Tissue adheres to the explant’s surface with help from an extracellular matrix (ECM), such as collagen or plasma clot. It can also happen by itself. Cells can migrate from the explant’s periphery. This is called a primary explant. Migrating cells are called outgrowth. This is used to compare the growth characteristics of cancer-cells to those of normal cells, particularly with regard to altered cell morphology and growth patterns.
Cell culture
This is the most common method for tissue culture. It involves the collection of cells that have grown from explants or dispersed suspensions (floating in culture medium). Cells that have been grown by mechanical or enzymatic means can be cultured on solid substrate as adherent monolayers.
Cell culture can be divided into three types: (1) Precursor cell culture, where undifferentiated cells are committed to differentiation; (2) Differentiated cell culture which is fully differentiated cells with lost the ability to differentiate further; and (3) Stem cell culture which is undifferentiated cell cultures that can develop into any kind of cell.
A cell line is a collection of cells that have a specific cell type and characteristic. These cells are then cloned or used in other ways to create a cell strain.
Monolayer cultures
Monolayer culture is an anchorage dependent culture that has one cell thickness and a continuous layer at the bottom.
Suspension Animal Cell Culture
Some cells can be kept mechanically suspended because they are not adhesive, which is a major advantage over monolayer cells (e.g. cells of leukemia). This has many advantages for the propagation and maintenance of cells.