Tissue culture

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Flasks containing tissue culture growth medium which provides nourishment for the growing of cells. Tissue culture vials nci-vol-2142-300.jpg
Flasks containing tissue culture growth medium which provides nourishment for the growing of cells.

Tissue culture is the growth of tissues or cells in an artificial medium separate from the parent organism. This technique is also called micropropagation. This is typically facilitated via use of a liquid, semi-solid, or solid growth medium, such as broth or agar. Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants. The term "tissue culture" was coined by American pathologist Montrose Thomas Burrows. [1] This is possible only in certain conditions. It also requires more attention. It can be done only in genetic labs with various chemicals.


Historical use

In 1885 Wilhelm Roux removed a section of the medullary plate of an embryonic chicken and maintained it in a warm saline solution for several days, establishing the basic principle of tissue culture. In 1907 the zoologist Ross Granville Harrison demonstrated the growth of frog embryonic cells that would give rise to nerve cells in a medium of clotted lymph. In 1913, E. Steinhardt, C. Israeli, and R. A. Lambert grew vaccinia virus in fragments of guinea pig corneal tissue. [2] In 1996, the first use of regenerative tissue was used to replace a small length of urethra, which led to the understanding that the technique of obtaining samples of tissue, growing it outside the body without a scaffold, and reapplying it, can be used for only small distances of less than 1 cm. [3]

Gottlieb Haberlandt first pointed out the possibilities of the culture of isolated tissues, plant tissue culture. [4] He suggested that the potentialities of individual cells via tissue culture as well as that the reciprocal influences of tissues on one another could be determined by this method. Since Haberlandt's original assertions, methods for tissue and cell culture have been realized, leading to significant discoveries in biology and medicine. His original idea, presented in 1902, was called totipotentiality: “Theoretically all plant cells are able to give rise to a complete plant.” [5] [6] [7]

Modern usage

Cultured cells growing in growth medium Cho cells adherend2.jpg
Cultured cells growing in growth medium

In modern usage, "Tissue culture" generally refers to the growth of cells from a multicellular organism in vitro. These cells may be cells isolated from a donor organism (primary cells) or an immortalised cell line. The cells are bathed in a culture medium, which contains essential nutrients and energy sources necessary for the cells' survival. [8] Thus, in its broader sense, "tissue culture" is often used interchangeably with "cell culture". On the other hand, the strict meaning of "tissue culture" refers to the culturing of tissue pieces, i.e. explant culture.

Tissue culture is an important tool for the study of the biology of cells from multicellular organisms. It provides an in vitro model of the tissue in a well defined environment which can be easily manipulated and analysed. In animal tissue culture, cells may be grown as two-dimensional monolayers (conventional culture) or within fibrous scaffolds or gels to attain more naturalistic three-dimensional tissue-like structures (3D culture). Eric Simon, in a 1988 NIH SBIR grant report, showed that electrospinning could be used to produced nano- and submicron-scale polymeric fibrous scaffolds specifically intended for use as in vitro cell and tissue substrates. This early use of electrospun fibrous lattices for cell culture and tissue engineering showed that various cell types would adhere to and proliferate upon polycarbonate fibers. It was noted that as opposed to the flattened morphology typically seen in 2D culture, cells grown on the electrospun fibers exhibited a more rounded 3-dimensional morphology generally observed of tissues in vivo. [9]

Plant tissue culture in particular is concerned with the growing of entire plants from small pieces of plant tissue, cultured in medium. [10] The technique of plant tissue culture, i.e., culturing plant cells or tissues in artificial medium supplemented with required nutrients, has many applications in efficient clonal propagation (true to the type or similar) which may be difficult via conventional breeding methods. Tissue culture is used in creating genetically modified plants, as it allows scientists to introduce DNA changes to plant tissue via Agrobacterium tumefaciens or a gene gun and then generate a full plant from these modified cells. [11]

Because plant cells are totipotent, adding growth hormones to the media can trigger the callus cells to develop roots, shoots and entire plants. [12]

Animal tissue culture

There are three common methods to establish cell culture from animals. The first is organ culture where whole organs from embryos or partial adult organs are used to initiate the organ culture in vitro. These cells retain their differentiated character and functional activity in organ culture. The second method is primary explant culture, in which fragments derived from animal tissue are attached to a surface using an extracellular matrix component (ECM), such as collagen or a plasma clot. This culture is known as a primary explant, and migrating cells are known as outgrowth. This has been used to analyze the growth characteristics of cancer cells in comparison to their normal counterparts. [13] The third method is cell culture, of which there are three types: (1) precursor cell culture, i.e. undifferentiated cells that are to be differentiate, (2) differentiated cell culture, i.e. completely differentiated cells that have lost the capacity to further differentiate, and (3) stem cell culture, i.e. undifferentiated cells that can develop into any kind of cell.” [13]

Applications of animal cell culture

Animal cell culture is used for many research purposes and commercial business also as:

Establishing a cell line

A cell line can be defined as a permanently established cell culture which will propagate forever. Investigators mostly get cell lines from other investigators or from the cell bank as The (American Type Culture Collection) , because its much easier than creating new one but in special cases investigators are obligated to establish a cell line to do this you must use one of the following cells:

Transformed cell lines, Tumor tissue or Transforming normal cell in vitro [14]


Subculture is the transfer of cells from one culture to start a new one. During this process the proliferating cells are subdivided, to form a new cell lines. [15]

Stem cell technology

The most advanced tissue culture science is now focused on stem cells, stem cells can be used for tissue replacement or either organs. stem cell is a primitive type of cell which has the ability to differentiate to all the 220 cell types found in human body. Stem cells can be obtained from blood, brain, or muscle tissue but the most important one is from early embryos which has the capability to differentiate to any other cell. [14]

See also

Related Research Articles

<span class="mw-page-title-main">Tissue (biology)</span> Group of cells having similar appearance and performing the same function

In biology, tissue is a historically derived biological organizational level between cells and a complete organ. A tissue is therefore often thought of as an assembly of similar cells and their extracellular matrix from the same origin that together carry out a specific function. Organs are then formed by the functional grouping together of multiple tissues.

<span class="mw-page-title-main">Tissue engineering</span> Biomedical engineering discipline

Tissue engineering is a biomedical engineering discipline that uses a combination of cells, engineering, materials methods, and suitable biochemical and physicochemical factors to restore, maintain, improve, or replace different types of biological tissues. Tissue engineering often involves the use of cells placed on tissue scaffolds in the formation of new viable tissue for a medical purpose but is not limited to applications involving cells and tissue scaffolds. While it was once categorized as a sub-field of biomaterials, having grown in scope and importance it can be considered as a field of its own.

Organ culture is the cultivation of either whole organs or parts of organs in vitro. It is a development from tissue culture methods of research, as the use of the actual in vitro organ itself allows for more accurate modelling of the functions of an organ in various states and conditions.

<span class="mw-page-title-main">Bioreactor</span> System that supports a biologically active environment

A bioreactor refers to any manufactured device or system that supports a biologically active environment. In one case, a bioreactor is a vessel in which a chemical process is carried out which involves organisms or biochemically active substances derived from such organisms. This process can either be aerobic or anaerobic. These bioreactors are commonly cylindrical, ranging in size from litres to cubic metres, and are often made of stainless steel. It may also refer to a device or system designed to grow cells or tissues in the context of cell culture. These devices are being developed for use in tissue engineering or biochemical/bioprocess engineering.

<span class="mw-page-title-main">Callus (cell biology)</span> Growing mass of unorganized plant parenchyma cells

Plant callus is a growing mass of unorganized plant parenchyma cells. In living plants, callus cells are those cells that cover a plant wound. In biological research and biotechnology callus formation is induced from plant tissue samples (explants) after surface sterilization and plating onto tissue culture medium in vitro. The culture medium is supplemented with plant growth regulators, such as auxin, cytokinin, and gibberellin, to initiate callus formation or somatic embryogenesis. Callus initiation has been described for all major groups of land plants.

<span class="mw-page-title-main">Cell culture</span> Process by which cells are grown under controlled conditions

Cell culture or tissue culture is the process by which cells are grown under controlled conditions, generally outside of their natural environment. The term "tissue culture" was coined by American pathologist Montrose Thomas Burrows. This technique is also called micropropagation. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. They need to be kept at body temperature (37 °C) in an incubator. These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or rich medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases (CO2, O2), and regulates the physio-chemical environment (pH buffer, osmotic pressure, temperature). Most cells require a surface or an artificial substrate to form an adherent culture as a monolayer (one single-cell thick), whereas others can be grown free floating in a medium as a suspension culture. This is typically facilitated via use of a liquid, semi-solid, or solid growth medium, such as broth or agar. Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants. The lifespan of most cells is genetically determined, but some cell-culturing cells have been “transformed” into immortal cells which will reproduce indefinitely if the optimal conditions are provided.

Organogenesis is the phase of embryonic development that starts at the end of gastrulation and continues until birth. During organogenesis, the three germ layers formed from gastrulation form the internal organs of the organism.

<span class="mw-page-title-main">Micropropagation</span> Practice in plant tissue culture

Micropropagation or tissue culture is the practice of rapidly multiplying plant stock material to produce many progeny plants, using modern plant tissue culture methods.

In biology, explant culture is a technique to organotypically culture cells from a piece or pieces of tissue or organ removed from a plant or animal. The term explant can be applied to samples obtained from any part of the organism. The extraction process is extensively sterilized, and the culture can be typically used for two to three weeks.

In biology, a substrate is the surface on which an organism lives. A substrate can include biotic or abiotic materials and animals. For example, encrusting algae that lives on a rock can be itself a substrate for an animal that lives on top of the algae. Inert substrates are used as growing support materials in the hydroponic cultivation of plants. In biology substrates are often activated by the nanoscopic process of substrate presentation.

Neural tissue engineering is a specific sub-field of tissue engineering. Neural tissue engineering is primarily a search for strategies to eliminate inflammation and fibrosis upon implantation of foreign substances. Often foreign substances in the form of grafts and scaffolds are implanted to promote nerve regeneration and to repair damage caused to nerves of both the central nervous system (CNS) and peripheral nervous system (PNS) by an injury.

A nerve guidance conduit is an artificial means of guiding axonal regrowth to facilitate nerve regeneration and is one of several clinical treatments for nerve injuries. When direct suturing of the two stumps of a severed nerve cannot be accomplished without tension, the standard clinical treatment for peripheral nerve injuries is autologous nerve grafting. Due to the limited availability of donor tissue and functional recovery in autologous nerve grafting, neural tissue engineering research has focused on the development of bioartificial nerve guidance conduits as an alternative treatment, especially for large defects. Similar techniques are also being explored for nerve repair in the spinal cord but nerve regeneration in the central nervous system poses a greater challenge because its axons do not regenerate appreciably in their native environment.

<span class="mw-page-title-main">Plant tissue culture</span> Growing plant cells under known conditions

Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues, or organs under sterile conditions on a nutrient culture medium of known composition. It is widely used, to produce clones of a plant in a method known as micropropagation. Different techniques in plant tissue culture may offer certain advantages over traditional methods of propagation, including:

Nano-scaffolding or nanoscaffolding is a medical process used to regrow tissue and bone, including limbs and organs. The nano-scaffold is a three-dimensional structure composed of polymer fibers very small that are scaled from a Nanometer scale. Developed by the American military, the medical technology uses a microscopic apparatus made of fine polymer fibers called a scaffold. Damaged cells grip to the scaffold and begin to rebuild missing bone and tissue through tiny holes in the scaffold. As tissue grows, the scaffold is absorbed into the body and disappears completely.

Tissue engineering of oral mucosa combines cells, materials and engineering to produce a three-dimensional reconstruction of oral mucosa. It is meant to simulate the real anatomical structure and function of oral mucosa. Tissue engineered oral mucosa shows promise for clinical use, such as the replacement of soft tissue defects in the oral cavity. These defects can be divided into two major categories: the gingival recessions which are tooth-related defects, and the non tooth-related defects. Non tooth-related defects can be the result of trauma, chronic infection or defects caused by tumor resection or ablation. Common approaches for replacing damaged oral mucosa are the use of autologous grafts and cultured epithelial sheets.

<span class="mw-page-title-main">Decellularization</span>

Decellularization is the process used in biomedical engineering to isolate the extracellular matrix (ECM) of a tissue from its inhabiting cells, leaving an ECM scaffold of the original tissue, which can be used in artificial organ and tissue regeneration. Organ and tissue transplantation treat a variety of medical problems, ranging from end organ failure to cosmetic surgery. One of the greatest limitations to organ transplantation derives from organ rejection caused by antibodies of the transplant recipient reacting to donor antigens on cell surfaces within the donor organ. Because of unfavorable immune responses, transplant patients suffer a lifetime taking immunosuppressing medication. Stephen F. Badylak pioneered the process of decellularization at the McGowan Institute for Regenerative Medicine at the University of Pittsburgh. This process creates a natural biomaterial to act as a scaffold for cell growth, differentiation and tissue development. By recellularizing an ECM scaffold with a patient’s own cells, the adverse immune response is eliminated. Nowadays, commercially available ECM scaffolds are available for a wide variety of tissue engineering. Using peracetic acid to decellularize ECM scaffolds have been found to be false and only disinfects the tissue.

A 3D cell culture is an artificially created environment in which biological cells are permitted to grow or interact with their surroundings in all three dimensions. Unlike 2D environments, a 3D cell culture allows cells in vitro to grow in all directions, similar to how they would in vivo. These three-dimensional cultures are usually grown in bioreactors, small capsules in which the cells can grow into spheroids, or 3D cell colonies. Approximately 300 spheroids are usually cultured per bioreactor.

<span class="mw-page-title-main">Margaret Reed Lewis</span> American cell biologist

Margaret Adaline Reed Lewis (1881–1970) was an American cell biologist and embryologist who made contributions to cancer research and cell culture techniques, and was likely the first person to successfully grow mammalian tissue in vitro. She authored around 150 papers, many co-authored with her husband Warren Harmon Lewis. The Lewises developed a growth medium called the Locke-Lewis solution and jointly received the Gerhard Gold Medal from the Pathological Society of Philadelphia.

In vitro spermatogenesis is the process of creating male gametes (spermatozoa) outside of the body in a culture system. The process could be useful for fertility preservation, infertility treatment and may further develop the understanding of spermatogenesis at the cellular and molecular level. 

Entomoculture is the subfield of cellular agriculture which specifically deals with the production of insect tissue in vitro. It draws on principles more generally used in tissue engineering and has scientific similarities to Baculovirus Expression Vectors or soft robotics. The field has mainly been proposed because of its potential technical advantages over mammalian cells in generating cultivated meat. The name of the field was coined by Natalie Rubio at Tufts University.


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