tissue culture

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 anspecific term plant tissue culture is used for plants.product cells have the same genotype (unless affected by mutation during culture)

Historical usageIn 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 usageIn 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 requiremen1. 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 semisolid3. Aseptic (sterile) conditions, as microorganisms grow much

animal tissue and can over run a culture4. 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 interest5. 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 removeda 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 requiremenAppropriate 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 semisolidAseptic (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


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

ClassificationTissue culture may be of two types based on medium.

1. Plant tissue culture2. 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 plantother 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 fre) 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 givplant 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


two types based on medium.Plant tissue cultureAnimal 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


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

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 needed1. A single explant can be multiplied into several thousand plants in less than a year

commercial propagation of new cultivars2. Taking an explant does not usually destroy the mother plant, so rare and endangered plants can be

cloned safely3. Once established, a plant tissue culture line can give a continuous supply of young plants throughout

the year4. In plants prone to virus diseases, virus free explants (new meristem tissue is usually virus free) can

be cultivated to provide virus free plants5. Plant ‘tissue banks’ can be frozen, then regenerated through tissue culture6. 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 cloned8. Tissue culture clones are ‘true to type’ as compared with seedlings, which show greater variability

Applications of Plant Tissue CulturePlant 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 speciesb) 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).

https://www.facebook.com/syed.m.dewanmass 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 neededcan be multiplied into several thousand plants in less than a year

commercial propagation of new cultivarsTaking 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 culturePlant 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 Cultureculture 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).


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

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 shortregeneration 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 CultureThe 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 intothe 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 medideveloped and the techniques of cell culture were refined.

Culturing Animal Tissues- The steps1. Animal tissue is obtained either from a particular

specimen, or from a ‘tissue bank’ of cryo(cryo = frozen at very low temperatures in a specimedium)

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 culture1. Animal cell lines have limited numbers of cell cycles before they begin to degrade2. Animal cells need frequent subculturing to remain viable

https://www.facebook.com/syed.m.dewanFor 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 dispersedcells 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 intothe 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 stepsAnimal 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 requiredHuman cells, for example are grown at 37degrees

differences from plant tissue cultureAnimal cell lines have limited numbers of cell cycles before they begin to degradeAnimal cells need frequent subculturing to remain viable

https://www.facebook.com/syed.m.dewanFor 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 intothe 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


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 Culture1. Growing viruses - these require

living host cells2. Making monoclonal antibodies, used

for diagnosis and research3. Studying basic cell processes4. Genetic modification & analysis5. ‘Knockout’ technology -

inactivating certain genes and tracing their effects

6. Providing DNA for the Human Genome Project (and other species’ genome projects)

Bibliography1. Dodds, J.H., Roberts, L.W., 1995, Experiments in Plant Tissue Culture, 3rd ed., Cambridge

University Press2. Hartmann, H., Kester, D., et.al., 1997, Plant Propagation, 6th ed., Prentice Hall International3. http://www.une.edu.au/agronomy/AgSrHortTCinfo.html4. http://aggie-horticulture.tamu.edu /tisscult/pltissue/pltissue.html5. http://www.liv.ac.uk/~sd21/tisscult/what.htm6. http://user.school.net.th/~anoparp/bptc1.htm

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