Tissue culture involves growing tissues or cells separate from the original organism. There are two main types - plant tissue culture and animal tissue culture. Plant tissue culture is used to propagate plants using techniques like micropropagation that offer advantages over traditional propagation methods. It relies on the totipotency of plant cells. Animal tissue culture refers to culturing whole organs, tissues or dispersed cells in a nutrient medium and is used in research applications.

1. Tissue Culture
Tissue culture is the growth of tissues and/or cells separate from the organism. This is
typically facilitated via use of a liquid, semi
Tissue culture commonly refers to the culture of animal cells an
specific term plant tissue culture is used for plants.
product cells have the same genotype (unless affected by mutation during culture)
Historical usage
In 1885 Wilhelm Roux removed
a portion 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 nerve cell processes in a
medium of clotted lymph.
Modern usage
In modern usage, "tissue culture" generally
refers to the growth of eukaryotic cells in vitro. It is often used interchangeably with cell culture to
specifically describe the in vitro culturing
be used to refer to the culturing of tissue pieces, i.e. explant culture or whole organs, i.e. organ
culture. It is a tool for the study of animal cell biology in vitro model of cell growth to allow
highly selective environment which is easily manipulated (used to optimize cell signaling
pathways). In 1920 germen scientist Harbland is considered as the father of tissue culture.
Tissue culture, both plant and animal has several critical requiremen
1. Appropriate tissue (some tissues culture better than others)
2. A suitable growth medium containing energy sources and inorganic salts to supply cell
growth needs. This can be liquid or semisolid
3. Aseptic (sterile) conditions, as microorganisms grow much
animal tissue and can over run a culture
4. Growth regulators - in plants, both auxins & cytokinins. In animals, this is not as well
defined and the growth substances are provided in serum from the cell types of interest
5. Frequent sub culturing to ensure adequate nutrition and to avoid the
metabolites
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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. Tissue culture produces clones, in which all
product cells have the same genotype (unless affected by mutation during culture)
In 1885 Wilhelm Roux removed
a portion 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
cell processes in a
In modern usage, "tissue culture" generally
refers to the growth of eukaryotic cells in vitro. It is often used interchangeably with cell culture to
specifically describe the in vitro culturing of sperm donor cells. However, "tissue culture" can also
be used to refer to the culturing of tissue pieces, i.e. explant culture or whole organs, i.e. organ
culture. It is a tool for the study of animal cell biology in vitro model of cell growth to allow
highly selective environment which is easily manipulated (used to optimize cell signaling
pathways). In 1920 germen scientist Harbland is considered as the father of tissue culture.
Tissue culture, both plant and animal has several critical requiremen
Appropriate tissue (some tissues culture better than others)
A suitable growth medium containing energy sources and inorganic salts to supply cell
growth needs. This can be liquid or semisolid
Aseptic (sterile) conditions, as microorganisms grow much more quickly than plant and
animal tissue and can over run a culture
in plants, both auxins & cytokinins. In animals, this is not as well
defined and the growth substances are provided in serum from the cell types of interest
to ensure adequate nutrition and to avoid the buildup
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Tissue culture is the growth of tissues and/or cells separate from the organism. This is
solid, or solid growth medium, such as broth or agar.
d tissues, while the more
Tissue culture produces clones, in which all
refers to the growth of eukaryotic cells in vitro. It is often used interchangeably with cell culture to
of sperm donor cells. However, "tissue culture" can also
be used to refer to the culturing of tissue pieces, i.e. explant culture or whole organs, i.e. organ
culture. It is a tool for the study of animal cell biology in vitro model of cell growth to allow a
highly selective environment which is easily manipulated (used to optimize cell signaling
pathways). In 1920 germen scientist Harbland is considered as the father of tissue culture.
Tissue culture, both plant and animal has several critical requirements
A suitable growth medium containing energy sources and inorganic salts to supply cell
more quickly than plant and
in plants, both auxins & cytokinins. In animals, this is not as well
defined and the growth substances are provided in serum from the cell types of interest
buildup of waste

2. Classification
Tissue culture may be of two types based on medium.
1. Plant tissue culture
2. Animal tissue culture
Plant Tissue Culture
Plant tissue culture is a practice used to propagate plants under sterile conditions, often to produce
clones of a plant. Different techniques in plant tissue culture may offer certain advantages over traditional
methods of propagation, including:
a) The production of exact copies of plant
other desirable traits.
b) To quickly produce mature plants.
c) The production of multiples of plants in the absence of seeds or necessary pollinators to produce
seeds.
d) The regeneration of whole plants fr
e) The production of plants in sterile containers that allows them to be moved with greatly reduced
chances of transmitting diseases, pests, and pathogens.
f) The production of plants from seeds that otherwise
growing, i.e.: orchids and nepenthes.
g) To clean particular plant of viral and other infections and to quickly multiply these plants as
'cleaned stock' for horticulture and agriculture.
Plant tissue culture relies on the fact that many plant cells have the ability to regenerate a whole
plant (totipotency). Single cells, plant cells without cell walls (protoplasts), pieces of leaves, or (less
commonly) roots can often be used to generate a new plant on culture media giv
plant hormones.
Culturing (micro propagating) Plant Tissue
1. Selection of the plant tissue (explant) from a
healthy vigorous ‘mother plant’
apical bud, but can be other tissue
2. This tissue must be sterilized to remove microbial
contaminants
3. Establishment of the explant in a culture medium.
The medium sustains the plant cells and
encourages cell division. It can be solid or liquid
4. Each plant species (and sometimes the variety
within a species) has particular medium
requirements that must be established by trial and
error
5. Multiplication- The explant gives rise to a callus (a
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two types based on medium.
Plant tissue culture
Animal tissue culture
ice used to propagate plants under sterile conditions, often to produce
clones of a plant. Different techniques in plant tissue culture may offer certain advantages over traditional
The production of exact copies of plants that produce particularly good flowers, fruits, or have
To quickly produce mature plants.
The production of multiples of plants in the absence of seeds or necessary pollinators to produce
The regeneration of whole plants from plant cells that have been genetically modified.
The production of plants in sterile containers that allows them to be moved with greatly reduced
chances of transmitting diseases, pests, and pathogens.
The production of plants from seeds that otherwise have very low chances of germinating and
growing, i.e.: orchids and nepenthes.
To clean particular plant of viral and other infections and to quickly multiply these plants as
'cleaned stock' for horticulture and agriculture.
the fact that many plant cells have the ability to regenerate a whole
plant (totipotency). Single cells, plant cells without cell walls (protoplasts), pieces of leaves, or (less
commonly) roots can often be used to generate a new plant on culture media given the required nutrients and
Culturing (micro propagating) Plant Tissue - the steps
Selection of the plant tissue (explant) from a
healthy vigorous ‘mother plant’ - this is often the
apical bud, but can be other tissue
must be sterilized to remove microbial
Establishment of the explant in a culture medium.
The medium sustains the plant cells and
encourages cell division. It can be solid or liquid
Each plant species (and sometimes the variety
ecies) has particular medium
requirements that must be established by trial and
The explant gives rise to a callus (a
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ice used to propagate plants under sterile conditions, often to produce
clones of a plant. Different techniques in plant tissue culture may offer certain advantages over traditional
s that produce particularly good flowers, fruits, or have
The production of multiples of plants in the absence of seeds or necessary pollinators to produce
om plant cells that have been genetically modified.
The production of plants in sterile containers that allows them to be moved with greatly reduced
have very low chances of germinating and
To clean particular plant of viral and other infections and to quickly multiply these plants as
the fact that many plant cells have the ability to regenerate a whole
plant (totipotency). Single cells, plant cells without cell walls (protoplasts), pieces of leaves, or (less
en the required nutrients and

3. mass of loosely arranged cells) which is
manipulated by varying sugar
concentrations and the auxin (low):
cytokinin (high) ratios to form multiple
shoots
6. The callus may be subdivided a number
of times
7. Root formation - The shoots are transferred to a growth medium with relatively higher auxin:
cytokinin ratios
8. The rooted shoots are potted up (deflasked) and
9. This is necessary as many young tissue culture plants have no waxy cuticle to prevent water loss
Why plant tissue culture is needed
1. A single explant can be multiplied into several thousand plants in less than a year
commercial propagation of new cultivars
2. Taking an explant does not usually destroy the mother plant, so rare and endangered plants can be
cloned safely
3. Once established, a plant tissue culture line can give a continuous supply of young plants throughout
the year
4. In plants prone to virus diseases, virus free explants (new meristem tissue is usually virus free) can
be cultivated to provide virus free plants
5. Plant ‘tissue banks’ can be frozen, then regenerated through tissue culture
6. Plant cultures in approved media are easier to export than are soil
pathogen free and take up little space (most current plant export is now done in this manner)
7. Tissue culture allows fast selection for crop improvement
then cloned
8. Tissue culture clones are ‘true to type’ as compared with seedlings, which show greater variability
Applications of Plant Tissue Culture
Plant tissue culture is used widely in plant science; it also has a number of commercial applications.
Applications include:
a) Micropropagation is widely used in forestry and in floriculture. Micropropagation can also be used
to conserve rare or endangered plant species
b) A plant breeder may use tissue culture to screen cells rather than plants for advantageous characters,
e.g. herbicide resistance/tolerance.
c) Large-scale growth of plant cells in liquid culture inside bioreactors as a source of secondary
products, like recombinant proteins used as biopharmaceuticals.
d) To cross distantly related species by protoplast fusion and regeneration of the novel hybrid.
e) To cross-pollinate distantly related species and then tissue culture the resulting embryo which would
otherwise normally die (Embryo Rescue).
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mass of loosely arranged cells) which is
manipulated by varying sugar
concentrations and the auxin (low):
okinin (high) ratios to form multiple
The callus may be subdivided a number
The shoots are transferred to a growth medium with relatively higher auxin:
The rooted shoots are potted up (deflasked) and ‘hardened off’ by gradually decreasing the humidity
This is necessary as many young tissue culture plants have no waxy cuticle to prevent water loss
Why plant tissue culture is needed
can be multiplied into several thousand plants in less than a year
commercial propagation of new cultivars
Taking an explant does not usually destroy the mother plant, so rare and endangered plants can be
a plant tissue culture line can give a continuous supply of young plants throughout
In plants prone to virus diseases, virus free explants (new meristem tissue is usually virus free) can
be cultivated to provide virus free plants
nks’ can be frozen, then regenerated through tissue culture
Plant cultures in approved media are easier to export than are soil-grown plants, as they are
pathogen free and take up little space (most current plant export is now done in this manner)
ulture allows fast selection for crop improvement - explants are chosen from superior plants,
Tissue culture clones are ‘true to type’ as compared with seedlings, which show greater variability
Applications of Plant Tissue Culture
culture is used widely in plant science; it also has a number of commercial applications.
Micropropagation is widely used in forestry and in floriculture. Micropropagation can also be used
to conserve rare or endangered plant species.
A plant breeder may use tissue culture to screen cells rather than plants for advantageous characters,
e.g. herbicide resistance/tolerance.
scale growth of plant cells in liquid culture inside bioreactors as a source of secondary
ombinant proteins used as biopharmaceuticals.
To cross distantly related species by protoplast fusion and regeneration of the novel hybrid.
pollinate distantly related species and then tissue culture the resulting embryo which would
ally die (Embryo Rescue).
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The shoots are transferred to a growth medium with relatively higher auxin:
‘hardened off’ by gradually decreasing the humidity
This is necessary as many young tissue culture plants have no waxy cuticle to prevent water loss
can be multiplied into several thousand plants in less than a year - this allows fast
Taking an explant does not usually destroy the mother plant, so rare and endangered plants can be
a plant tissue culture line can give a continuous supply of young plants throughout
In plants prone to virus diseases, virus free explants (new meristem tissue is usually virus free) can
grown plants, as they are
pathogen free and take up little space (most current plant export is now done in this manner)
explants are chosen from superior plants,
Tissue culture clones are ‘true to type’ as compared with seedlings, which show greater variability
culture is used widely in plant science; it also has a number of commercial applications.
Micropropagation is widely used in forestry and in floriculture. Micropropagation can also be used
A plant breeder may use tissue culture to screen cells rather than plants for advantageous characters,
scale growth of plant cells in liquid culture inside bioreactors as a source of secondary
To cross distantly related species by protoplast fusion and regeneration of the novel hybrid.
pollinate distantly related species and then tissue culture the resulting embryo which would

4. f) For production of doubled monoploid (dihaploid) plants from haploid cultures to achieve
homozygous lines more rapidly in breeding programmes, usually by treatment with colchicine
which causes doubling of the chromosome number.
g) As a tissue for transformation, followed by either short
regeneration of transgenic plants.
h) Certain techniques such as meristem tip culture can be used to produce clean plant material from
virused stock, such as potatoes
i) micropropagation using meristem and shoot culture to produce large numbers of identical
individuals
Animal Tissue Culture
The term tissue culture refers to the culture of whole organs, tissue fragments as well as dispersed
cells on a suitable nutrient medium. It can be divided into
organisation is retained or not.
The beginning of animal tissue culture can be traced to 1880 when Arnold showed t
can divide outside the body. Later, in 1903, Jolly studied the behaviour of animal tissue explants immersed
in serum, lymph or ascites fluid.
In 1907, Harrison cultured frog tadpole spinal chord in a lymph drop hanging from a cover slip into
the cavity on a microscopic slide; this is regarded as the turning point. A few years later, in 1913, Carrel
developed a complicated methodology for maintaining cultures free from contamination, especially by
bacteria.
Subsequently, suitable culture medi
developed and the techniques of cell culture were refined.
Culturing Animal Tissues- The steps
1. Animal tissue is obtained either from a particular
specimen, or from a ‘tissue bank’ of cryo
(cryo = frozen at very low temperatures in a speci
medium)
2. Establishment of the tissue is accomplished in the
required medium under aseptic conditions
3. Growing the cells / tissue requires an optimum
temperature, and subculturing when required
4. Human cells, for example are grown at 37degrees
and 5% CO2
Animal tissue/cell culture - differences from plant tissue culture
1. Animal cell lines have limited numbers of cell cycles before they begin to degrade
2. Animal cells need frequent subculturing to remain viable
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For production of doubled monoploid (dihaploid) plants from haploid cultures to achieve
homozygous lines more rapidly in breeding programmes, usually by treatment with colchicine
which causes doubling of the chromosome number.
a tissue for transformation, followed by either short-term testing of genetic constructs or
regeneration of transgenic plants.
Certain techniques such as meristem tip culture can be used to produce clean plant material from
virused stock, such as potatoes and many species of soft fruit.
micropropagation using meristem and shoot culture to produce large numbers of identical
The term tissue culture refers to the culture of whole organs, tissue fragments as well as dispersed
cells on a suitable nutrient medium. It can be divided into
(1) organ culture and
(2) cell culture mainly on the basis of whether the tissue
organisation is retained or not.
The beginning of animal tissue culture can be traced to 1880 when Arnold showed t
can divide outside the body. Later, in 1903, Jolly studied the behaviour of animal tissue explants immersed
In 1907, Harrison cultured frog tadpole spinal chord in a lymph drop hanging from a cover slip into
the cavity on a microscopic slide; this is regarded as the turning point. A few years later, in 1913, Carrel
developed a complicated methodology for maintaining cultures free from contamination, especially by
Subsequently, suitable culture media were
developed and the techniques of cell culture were refined.
The steps
Animal tissue is obtained either from a particular
specimen, or from a ‘tissue bank’ of cryo-preserved
(cryo = frozen at very low temperatures in a special
Establishment of the tissue is accomplished in the
required medium under aseptic conditions
Growing the cells / tissue requires an optimum
temperature, and subculturing when required
Human cells, for example are grown at 37degrees
differences from plant tissue culture
Animal cell lines have limited numbers of cell cycles before they begin to degrade
Animal cells need frequent subculturing to remain viable
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For production of doubled monoploid (dihaploid) plants from haploid cultures to achieve
homozygous lines more rapidly in breeding programmes, usually by treatment with colchicine
term testing of genetic constructs or
Certain techniques such as meristem tip culture can be used to produce clean plant material from
micropropagation using meristem and shoot culture to produce large numbers of identical
The term tissue culture refers to the culture of whole organs, tissue fragments as well as dispersed
(2) cell culture mainly on the basis of whether the tissue
The beginning of animal tissue culture can be traced to 1880 when Arnold showed that leucocytes
can divide outside the body. Later, in 1903, Jolly studied the behaviour of animal tissue explants immersed
In 1907, Harrison cultured frog tadpole spinal chord in a lymph drop hanging from a cover slip into
the cavity on a microscopic slide; this is regarded as the turning point. A few years later, in 1913, Carrel
developed a complicated methodology for maintaining cultures free from contamination, especially by

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3. Tissue culture media is not as fully defined as that of plants - in addition to inorganic salts, energy
sources, amino acids, vitamins, etc., they require the addition of serum (bovine serum is very
common, but others are used)
4. Animal tissue cultures can pose biohazard concerns, and cultures require special inactivation with
hypochlorite (e.g. Janola,Chlorox,
etc.) and then incineration
Uses of Animal Tissue Culture
1. Growing viruses - these require
living host cells
2. Making monoclonal antibodies, used
for diagnosis and research
3. Studying basic cell processes
4. Genetic modification & analysis
5. ‘Knockout’ technology -
inactivating certain genes and
tracing their effects
6. Providing DNA for the Human
Genome Project (and other species’
genome projects)
Bibliography
1. Dodds, J.H., Roberts, L.W., 1995, Experiments in Plant Tissue Culture, 3rd ed., Cambridge
University Press
2. Hartmann, H., Kester, D., et.al., 1997, Plant Propagation, 6th ed., Prentice Hall International
3. http://www.une.edu.au/agronomy/AgSrHortTCinfo.html
4. http://aggie-horticulture.tamu.edu /tisscult/pltissue/pltissue.html
5. http://www.liv.ac.uk/~sd21/tisscult/what.htm
6. http://user.school.net.th/~anoparp/bptc1.htm