2. • Introduction
• Tissue Culture in Animals
• Types of Animal Cell Culture
• Summary Of Animal Tissue Culture
• Tissue Culture in Plants
• Types of Plant Cell Culture
• Explanations of the Types
• Major steps of plant tissue culture
• Technique for plant in vitro culture
• Conclusions
• References
Contents
3. Tissue Culture:
In biological research, tissues culture
refers to a method in which fragments of a plant or animal tissue
are introduced into a new, artificial environment, where they
continue to function or grow.
While fragments of a tissue are often used, it is important to note
that entire organs are also used for tissue culture purposes. Here,
such growth media as broth and agar are used to facilitate the
process.
Introduction
While the term tissue
culture may be used
for the both plant and
animals tissues, plant
tissue culture is the
more specific term
used for the culture of
plant tissues in tissue
culture.
4. • Tissue culture (TC) is the cultivation of plant cells, tissues, or
organs on specially formulated nutrient media. Under the right
conditions, an entire plant can be regenerated from a single
cell. Plant tissue culture is a technique that has been around
for more than 30 years. Tissue culture is seen as an important
technology for developing countries for the production of
disease-free, high quality planting material and the rapid
production of many uniform plants.
What is tissue Culture?
6. Summary of Animal Tissue Culture
Animal cell culture technology in today’s scenario has become
indispensable in the field of life sciences, which provides a basis
to study regulation, proliferation, and differentiation and to
perform genetic manipulation.
It requires specific technical skills to carry out successfully. This
chapter describes the essential techniques of animal cell culture
as well as its applications.
Animal cell culture is important to all fields of bioscience;
especially from medicine to agriculture. It is an important tool to
study cell specific functions, physiology and biochemical
components. The major advantage of cell culture is its
consistency and reproducibility of results that can be obtained
from using clonal cells. They serve as building blocks and stem
cells for bioscience research and biological repairs. Priority
should be given to developing good cell culture laboratories
especially in developing countries where they are not yet fully
established.
7. Tissue culture is applied in plant research for such purposes as the
growing of new plants, which in some cases undergo genetic
alterations.
Here, the plant of interest is taken through the tissue culture process
and grown in a controlled environment.
Representative images of stages in tissue culture from seeds to
plants. Inoculated seeds of I. sanguine from (A) sterile
indehiscent capsule and soaked in NaOH, or (B) hot water; (C)
germinated seeds and (D) seedlings; (E) induced adventitious
shoots at 20 and (F) 40 days; (G) clustered shoots with induced
roots (noticeable from the bottom of the jar); (H) transplanted
plantlets established after 30 days.
Plants tissue culture:
9. Meristem Culture:
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Shoot meristems, whether apical or axillary, are small domes
(50-150,um in diameter) consisting of groups of a few actively
dividing cells protected by developing leaves. The main
physiological functions of meristems are the synthesis of
protoplasm and the production of new cells. The biology and
developmental morphology of the shoot apical meristems have
been studied in grasses
Apical Meristem from angiosperm and gymnosperm shorts are
cultures to produce plants that are largely free from disease and
contamination.
10. Callus Culture:
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This is the term used to refer to unspecialized, unorganized and a
dividing mass of cells. A callus is produced when explants are cultured
in an appropriate medium.
A good example of this is the tumor tissue that grows out of the
wounds of differentiated tissues/organs.
In practice, callus culture involves the growth of callus which is the
followed by a procedure that induces organ differentiation for this
culture, the culture is often sustained on a gel medium, which is
composed of agar and a mixture of given macro and micronutrients
depending on the type of cells.
Callus Culture Development:
11. Seed culture:
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•Seed culture is the type of tissue culture that is primarily used
for plants such as orchids.
•For this method, explants are obtained from an in-vitro derived
plant and introduced in to an artificial environment, where they
get to proliferated.
•In Vitro Seed Culture Development:
12. Embryo Culture:
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Embryo culture is the type of tissue culture that involves in the
isolation of an embryo from a given organism for in-vitro growth.
The term embryo culture is used to refer to sexually produced zygotic
embryo culture.
Embryo culture may involve the use of a mature of immature embryo.
Whereas mature embryos for culture are essentially obtained from
ripe seed, immature embryo involves the use of immature embryo
from unripe/hybrid seeds that failed to germinate.
Embryo Culture Development :
13. Organ Culture:
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Organ culture is a type of tissue culture that involves isolating an
organ for in-vitro growth. Here, any organ plant can be used as an
explants for the culture process.
With organ culture, or as is with their various tissue components,
the method is used for preserve their structure or functions, which
allows the organ to still resemble and retain the characteristics they
would have in vivo.
There are number of methods that can be used for organ culture.
Plasma clot method (2) Raft method (3) Agar gel method (4) Grid
method
Diagram
14. Protoplast Culture:
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Protoplast- cells without cell walls. A protoplast is the term used to
refer to cell (fungi, bacteria, plant cell etc.) in which the cell wall has
been removed, which is why they are also referred to as hiked cell.
Protoplasts mat be cultured in the following ways.
(1)Hanging-drop culture
(2) Micro culture chambers
(3) Soft agars matrix
The following is a general
procedure for plant tissue
culture.
Medium preparation
Plant preparation
16. Stages:
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Initiation phase (Stage 1)
The initiation phase is the first phase of tissue culture. Here, the
tissue of interest is obtained and introduced and sterilized in
order to prevent any microorganism from negatively affecting
the process.
It is during this stage that the tissue is initiated in to culture.
Multiplication phase (Stage 2)
The multiplication phase is the 2nd step of tissue culture where
the in-vitro plant material is re-divided and then medium is
composed of appropriate components for growth including
regulators and nutrients.
These are responsible for the proliferation of the tissue and the
production of multiple shoots. This step is often repeated several
times in order to obtain the desired number of plant.
Root formation (Stage 3)
Its at this phase that roots are formed. Here, hormones are
required in order to induce rooting, and consequently complete
plantlets.
17. Technique for plant in vitro
culture
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Uses:
Micro propagation:
This technique is used for the purposes of developing high-
quality clonal plants. This has the potential to provide rapid
and large scale propagation of new genotypes.
Somatic cell genetics:
Used for haploid production and somatic hybridization.
Transgenic plants:
Used for expression of mammalian genes or plant genes for
various species it has proved beneficial for the engineering of
species that are resistant against viruses and insects.
18. Conclusion
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In reality, there are numerous methods used for tissue culture
give that there are different types of tissues that require specific
conditions for the culture process yield desired results.
both plant and animals tissue can be used for tissue culture
purposes for wide range of purposes for instance, animal tissue
culture may serve such purposes as preservation of an
organ/tissue studying the tutors or given tissues or for diagnosis
purposes on other hands, plant tissue culture may be used for
cloning purposes, genetic modification of a given plant or simple
to accelerate or increase yield of the plant of interest.
Tissue culture is therefore of great significance in biological
studies due to its wide range of application. The processes
involved in tissue culture may be complex, requiring a lot of care
to avoid such effects as contamination. Because of the
complexities that may be involved in some of the steps, this may
not be an experiment for everyone.