Tissue Culture

Group 7
Objectives
After this discussion, you will be able to:
Identify what is tissue culture.
Know the process/experiment of tissue culture and how it
works.
Identify the types of tissue culture.
Know the apparatus to be used.
Identify the precautionary measures.
Know the history , persons Involved , importance ,
application , advantages and disadvantages.
What is Tissue
Culture?
Tissue culture, a method of
biological research in which
fragments of tissue from an animal
or plant are transferred to an
artificial environment in which they
can continue to survive and
function.
Presentations
 Tissue culture is the technique of
growing cells and tissues in an
artificial medium separate from the
organism. This technique is also called
micropropogation.
Wikipedia, ( 2008)
History
 A German Botanist, made the first attempt
to use the in vitro method when grow plant
tissues. Although tissue culture has been
around since the beginning of the 18th
century, plant tissue culture only began
developing in 1898.

Persons Involved
 Gottlieb Haberlandt, a
German Botanist, made the
first attempt to use the in
vitro method when grow
plant tissues. The cells he
used were varied, palisade
tissues coming from:
-the leaves
-the piths
-the epidermis and
epidermal hairs
 An early attempt at tissue culture
was made in 1885 by German
zoologist Wilhelm Roux, who
cultivated tissue from a chick
embryo in a warm salt solution.

The first real success came in 1907,

however, when American zoologist

Ross G. Harrison demonstrated the

growth of frog nerve cell processes in
a medium of clotted lymph.
Apparatus to be used :
Safety equipment;
Laboratory coat
Gloves
Safety Goggles
First aid kit
Fire extinguisher
Ph meter -A pH meter is an instrument used to
measure hydrogen ion activity in solutions - in
other words, this instrument measures
acidity/alkalinity of a solution.
Volumetric flask-A volumetric flask is used
when it is necessary to know both precisely
and accurately the volume of the solution
that is being prepared.
 Pipette-A pipette is a laboratory
instrument used to measure out or transfer
small quantities of liquid.
Beaker-A beaker is a cylindrical glass or
plastic vessel used for holding liquids. It is
a multi- purpose piece of equipment used
for containing a chemical reaction and
measuring liquids.
Graduated Cylinders-used for measuring the
volumes of liquids.
One of the essential pieces of equipment
for any lab is a heat resistant Erlenmeyer
flask. These flasks will be used for
preparing the tissue culture media. Other
equipment pieces include a small capacity
flask and a culture jar.
 A Petri dish is a shallow transparent
lidded dish that biologists use to hold
growth medium in which cells can be
cultured
Types of Tissue Culture
 Seed Culture
In this culture, the explants are obtained from an
in-vitro derived plant and introduced into a
laboratory where they proliferate. The explant
should be sterilized to prevent it from tissue
damage.
 Protoplast Culture
It is a cell without a cell wall. A protoplast can be
cultured using the hanging-drop method, or
micro-culture chambers. In protoplast culture, a
number of phases can be observed: development
of cell wall, cell division, regeneration of a whole
plant.
 Embryo Culture
This involves the in-vitro development of an embryo.
For this, an embryo is isolated from a living organism.
Both, a mature or an immature embryo can be used in
the process. Mature embryos can be obtained from ripe
seeds. The immature embryos are obtained from the
seeds that failed to germinate. The ovule, seed or fruit
is already sterilized, therefore, it does not need to be
sterilized again.
 Callus Culture
A callus is an unorganized, dividing mass of
cells. When the explants are cultured in a
proper medium, the callus is obtained. The
growth of callus is followed by organ
differentiation. The culture is grown on a gel-
like medium composed of agar and specific
nutrients required for the growth of the cells.
 Organ Culture
In this, any organ of the plant such as shoot,
leaf, can be used as an explant. A number of
methods can be used for the organ culture,
such as plasma clot method, raft method, grid
method, and agar gel method. This method is
used to preserve the structure and functions of
an organism.
 Guidelines when working in a tissue culture lab:
1.Wear gloves and a lab coat and keep long hair tied
back.
2.Work in a laminar flow hood when passaging cells.
3.Wipe down working surfaces with ethanol.
4.Use sterile equipment.
 Stay as organized as possible—label everything
and set up all of your materials before getting
started.
Inspect all equipment and media for visible
contamination before use.
If you must completely remove a lid from a tube,
plate or bottle, set it down within the hood with
the open surface facing up. Otherwise, keep
tubes, plates or bottles closed as much as
possible.
 Do not pass your hands/arms over any open bottle,
plate or tube.
Use proper antibiotics in your culture media.
When finished, do the following: write observations,
methods and results in your lab notebook, put
everything away, dispose of materials properly, wipe
down working surfaces with ethanol, and turn on UV
lamp within laminar flow hood for 10 minutes to
sterilize the area.
 Importance
Tissue culture is a very important technology for
developing countries, for the production of disease-
free, high quality planting material and the rapid
production of many uniform plants , plant tissue
culture may be used for genetic modification of a
plant or simply increase it's yield.
 Application
• Tissue culture has given a mechanism for propagating
and genetically improving commercially valuable
plants.
• Tissue culture has been used to study characteristics of
the growth, metabolism, reproduction, physiology and
nutritional necessities of the plants under controlled
circumstances.
• Mutagens are added to single cell liquid cultures for
initiation of mutations
 • Embryos which normally do not survive inside seeds can be
developed in tissue culture to produce new plants. It can be used
for interspecific hybridization.

• Large scale fabrication of artificial seeds through somatic

embryogenesis.

• The plantlets are produced in a very short time with a lesser

amount of plant tissue.

• The production of multiples of plants in the lack of seeds or

essential pollinators to produce seeds.
Advantages of Tissue Culture
The new plantlets can be grown in a short amount of
time.
Only a small amount of initial plant tissue is required.
The new plantlets and plants are more likely to be
free of viruses and diseases.
The process is not dependant on the seasons and can
be done throughout the year.
The plantlets are obtained in a very short time with a
small amount of plant tissue.
The new plants produced are disease-free.

The plants can be grown throughout the year,

irrespective of the season.

A large space is not required to grow plants by tissue

culture technique.

The production of new varieties in the market place

speeds up.
This technique is being used
for the production of ornamental
plants such as dahlia, chrysanthemum, orchids, etc.
 Disadvantages

-Tissue Culture can require more labor

and cost more money.
-There is a chance that the propagated
plants will be less resilient to diseases
due to the type of environment they are
grown in.
It is imperative that, before being cultured, the
material is screened; failure to pick up any
abnormalities could lead to the new plants
being infected.
While the success rate is high if the correct
procedures are followed, success with the tissue
culture is not a guarantee. There is still a
chance that the process triggers a secondary
metabolic chemical reaction, and the new
explants or cells' growth gets stunted, or even
die off.
Short Quiz
 1.The technique of growing cells and
tissues in an artificial medium
separate from the organism.

2.A German Botanist, made the first

attempt to use the in vitro method when
grow plant tissues.
3. Give atleast one type of tissue
culture.
4.These flasks will be used for
preparing the tissue culture media.
5. Give atleast two safety equipments
that are used in the laboratory.
Thank you for
listening

Members:
Miera Monique Torrecampo
Era Jean Mecijas
Michelle Ann Jimenez
Ritchmond Dave Cano
Ethan James Magalona