Tissue culture

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Vials containing tissue culture growth medium which provides nourishment to growing cells.

Tissue culture is the growth of tissues and/or cells separate from the organism. 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, while the more specific term
plant tissue culture is used for plants.
 Cloning Plants:
Tissue Culture
Horticulture and the Science of Plants
Youth Adventure Program

David Wm. Reed, Instructor

Department of Horticultural Sciences
Texas A&M University

BACKGROUND

Tissue culture (often called micropropagation) is a special type of asexual propagation where a
very small piece of tissue (shoot apex, leaf section, or even an individual cell) is excised (cut-out)
and placed in sterile (aseptic) culture in a test tube, petri dish or tissue culture container containing a
special culture medium.

Overview of the Tissue Culture Process

The culture medium contains a gel (agar) with the proper mixture of nutrients, sugars, vitamins and
hormones, which causes the plant part to grow at very rapid rates to produce new plantlets. It has
been estimated that one chrysanthemum apex placed in tissue culture could produce up to 1,000,000
new plantlets in one year. Thus, tissue culture is used for rapid multiplication of plants. A very
specialized laboratory is required for tissue culture. All the procedures are done in a laboratory and
special ventilated cabinet that is as sterile as an operating room.

Steps in Tissue Culture

(images courtesy of Dr. Dan Lineberger, aggie-
horticulture.tamu.edu/tisscult/microprop/microprop.html)
Explant: Cut-out Plant Tissue and Place in Tissue Culture Container
The first step is to obtain what is called and explant. This means to simply cut-out a very small
piece of leaf or stem tissue, or even isolate individual cells, and place them in a tissue culture
container. The tissue has to be sterilized so it will not have any contaminating bacteria or fungus. It
is then placed inside the tissue culture contain on a gel called agar. In the agar is dissolved all the
sugar, nutrients and hormones the plant needs.

Explants can be pieces of any part of the plant (leaves, stems, flowers, etc.),
or even individual isolated cells.

Multiplication: Tissue Grows and Produces Small Plants

The tissue will begin to grow. It may make a big blob of tissue called callus, or it may make new
shoots directly from the explant tissue that was inserted in the container.

A mass of callus tissue is formed that is just starting to make new plantlets.
 New plantlets (shoots with leaves) are forming.

If the conditions are right a small "forest" of plants

will develop in the tissue culture container.

Rapid Multiplication by Transfer of Cultures

Once the plantlets start developing, some can be removed and placed in new tissue culture
containers. Thus, another "forest"' of plants is produced. This results in a rapid multiplication of the
cultures and many thousand of plants can be produced in a few months.
 Some of the small plantlets can be removed and transferred to new tissue culture
containers. These will produce more shoots and fill the container.

Transplanting
When the plantlets are large enough, they can be removed from the tissue culture container and
transferred into pots with potting soil. The young plants are growth in a greenhouse just like you
would any young seedling or cutting.

When the small plant clones are removed from the culture containers, they must be transplanted into
some type of acclimation container or kept under a mist system until the acclimate to the ambient
environment.
 After acclimation, the young plants can be transplanted
and grown in pots in a greenhouse to produce new plants.

TAKE HOME LESSONS

1) To become familiar with how plants can be cloned from very small pieces of leaf, stem or other
plant tissue.
2) To transfer plants out of a tissue culture container into a new tissue culture container.
3) Bring the new tissue culture clone home and transplant it into soil in a pot when it gets large
enough.

MATERIALS NEEDED
Plantlets in a tissue culture container ready to be transferred.
Tissue culture lab and transfer hood.
Tissue culture container with fresh tissue culture medium.

PROCEDURES
View a Video on Cloning with Tissue Culture and Observe a Variety of Tissue Cultures
The class go to a tissue culture lab and view a video on tissue culture.
The class will view a variety of tissue culture containers with cloned plantlets.

Transfer Cloned Plantlets to Fresh Tissue Culture Container

The class will be instructed on the proper procedure to:
The branch of biology in which tissues or cells of higher animals and plants are
grown artificially in a controlled environment. Tissue culture is possible when cells
are attached to a solid substrate, such as glass or cellophane, and if the necessary
complex nutrient medium is provided. All cultures are now also grown in liquid
suspension. Tissue cultures are used in the study of cell growth, multiplication, and
differentiation, as well as in cancer research, hereditary mechanisms, radiation
biology, all hybridization, and virus studies

tissue culture

Dictionary: tissue culture

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Home > Library > Literature & Language > Dictionary

n.

1. The technique or process of keeping tissue alive and growing in a culture medium.
2. A culture of tissue grown by this technique or process.

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5min Related Video: tissue culture

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Sci-Tech Encyclopedia: Tissue culture

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The branch of biology in which tissues or cells of higher animals and plants are grown artificially
in a controlled environment. Tissue culture is possible when cells are attached to a solid
substrate, such as glass or cellophane, and if the necessary complex nutrient medium is provided.
All cultures are now also grown in liquid suspension. Tissue cultures are used in the study of cell
growth, multiplication, and differentiation, as well as in cancer research, hereditary mechanisms,
radiation biology, all hybridization, and virus studies.
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Britannica Concise Encyclopedia: tissue culture
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Biological research method in which tissue fragments (a cell, a population of cells, or all or part
of an organ) are sustained in an artificial environment for examination and manipulation of cell
behaviour. It has been used to study normal and abnormal cell structure; biochemical, genetic,
and reproductive activity; metabolism, functions, and aging and healing processes; and reactions
to physical, chemical, and biological agents (e.g., drugs, viruses). A tiny sample of the tissue is
spread on or in a culture medium of biological (e.g., blood serum or tissue extract), synthetic, or
mixed origin having the appropriate nutrients, temperature, and pH for the cells being incubated.
The results are observed with a microscope, sometimes after treatment (e.g., staining) to
highlight particular features. Some viruses also grow in tissue cultures. Work with tissue cultures
has helped identify infections, enzyme deficiencies, and chromosomal abnormalities; classify
brain tumours; and formulate and test drugs and vaccines.

For more information on tissue culture, visit Britannica.com.

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Columbia Encyclopedia: tissue culture
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tissue culture, the propagation of plants through the placement of small amounts of
undifferentiated tissue or single cells in an artificial environment. The tissue is placed in a
nutrient medium that favors the production of roots and shoots, and is later planted normally. By
using tissue culture, the favorable qualities of plants can be precisely controlled, so that each
plant is identical for the particular quality being sought, whether it be disease resistance or plant
chemical production.

Gardener's Dictionary: tissue culture

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A laboratory technique of propagating new plants from tiny portions of the parent plant.