INTRODUCTION

Cell culture is the complex process by which cells are

grown under controlled conditions, generally outside of
their natural environment.
The cells can be derived from multi-cellular eukaryotes,
especially animal cells, plants, fungi and microbes,
including viruses, bacteria and protists.
CELL AND TISSUE CULTURE:
Tissue culture is the general term for the removal of cells, tissues and
organs from an animal or plant and their subsequent placement to the
artificial environment conductive to growth.
The culture of whole organs or intact organ fragments studying their
growth and function is known as organ culture.
When the cells are removed from the organ prior to or during cultivation,
thus disrupting their relationship with their neighboring cells is called as
cell culture.
METHODS TO OBTAIN CELL CULTURE:
When the cells from an organism are surgically removed and placed
into a suitable medium with controlled environment, they will attach to
the surface of the vessel, divide and grow. This is called primary
culture. This can be obtained by two methods.

 Mechanical disruption, the organ is divided in to small pieces around 1CC using
sterile sharp scalpel. Small pieces of tissue attached to the culture vessel in the
medium will move from the tissue explant and begin to divide and grow.
 Enzymatic dissociation is the second more widely used method which is
performed by adding proteolytic or digesting enzymes, such as trypsin or
collagenase that creates suspension of single cells that are then placed in to
culture vessels containing appropriate medium.
Subculturing:
When the cells in the primary culture reached confluency (grown and
filled up all the substrate available) they must be subcultured in to new
culture vessel containing fresh medium for their continued growth.
Subculturing is usually done by removing cells from the substrate as
gently as possible with enzymes or some cell lines harvested by gently
scraping the cells from the bottom.
Cell lines:
Cell line is a permanently established cells in a culture that will
proliferate indefinitely with supplementation of appropriate fresh
medium and space. In cell lines, cells become immortalized so it is also
known as infinite culture.
Finite cell Lines:
The cell lines which have a limited life span and go through a
limited number of cell generations approximately usually 20-80
population doublings are known as Finite cell lines. These cell lines
exhibit the property of contact inhibition, density limitation and
anchorage dependence. The growth rate is slow and doubling time
is around 24-96 hours.
Continuous Cell Lines:
Cell lines transformed under laboratory conditions or in vitro
culture conditions give rise to continuous cell lines. Absence of
contact inhibition and anchorage dependence.The growth rate is
rapid and doubling time is 12-24 hours..
CLASSIFICATION OF CELL LINES:
In terms of growth mode cells will grow either in suspension (as single
cells or small free floating clumps) or as a monolayer that is attached to
the tissue culture flask.
The form taken by a cell line reflects the tissue from which it was derived
e.g. cell lines derived from blood (leukaemia, lymphoma) tend to grow
in suspension whereas cells derived from solid tissue (lungs, kidney) tend
to grow as monolayers
Attached cell lines can be classified as endothelial epithelial HeLa,
neuronal and fibroblasts
Cells in culture can be divided in to three basic categories based
on their shape and appearance (i.e., morphology).
• Fibroblastic (or fibroblast-like) cells are bipolar or multipolar,
have elongated shapes, and grow attached to a substrate
Epithelial-like cells are polygonal in shape with more regular
dimensions, and grow attached to a substrate in discrete patches.
Lymphoblast-like cells are spherical in shape and usually grown in
suspension without attaching to a surface.
APPLICATIONS OF CELL CULTURE:
1. Model Systems: for studying Basic cell biology and Biochemistry, the
interaction between diseases causing agents and cells, the effects of
drugs on cell,etc.
2. Toxicity Testing: for studying the effect of new drugs, cosmetics and
chemicals on survival and growth in a wide variety of cell types.
3.Cancer research: to study the effect of chemicals used in chemotherapy
on normal cells and its efficiency on cancer cells as well as effect of
radiation during radiotherapy.
4. Virology: used in the clinical detection and isolation of virus ,as well as
for vaccine production
5. Cell based manufacturing: used for the production of many
important products like vaccines for polio and rabies,monoclonal
insulin, antibodies, hormones etc.
Tissue and organ transplantation therapy which include bone marrow
transplantation, adult and embryonic stem cell transplantation.
6. Genetic counselling: Amniocentesis enables doctors to diagnostic
foetal disorders in early stage of development. Cells are examined
for karyotyping, chromosome painting and other molecular
techniques.
7. Genetic engineering: Transfection and reprogramming the genetic
materials of the cultured cells is a major tool to molecular biology to
study the cellular effects of the expression of new proteins.
CULTURING HUMAN EMBRYONIC STEM CELLS
ADVANTAGES OF THE IN VITRO STUDIES:
Control of environment: Economy scale and mechanism:
Living organisms are extremely Cost of cell culture are far less
complex functional systems that compared to injecting in vivo. Screening
test with many replicates are cheaper,
are a great barrier to the legal, moral and ethical.
identification of individual
components and the exploration of
their basic biological functions. The
primary advantage of in vitro
work is that it permits an enormous
level of simplification of the
system under study, so that the
investigator can focus on a small
number of components.
LIMITATIONS OF IN VITRO STUDIES:
Animal cell culture needs expertise for handling techniques since it
needs strict aseptic conditions throughout the experiments.
Origin of cells: In tissue culture, a piece of tissue contains more
than one type of cells; when cells are differentiated in vitro there is
chance of losing original properties of cells. So that will be difficult
to relate to origin.
Instability: It has also been observed that, there is variation from
passage to passage and instability in cells also seen.
QUESTIONS
1. Define cell lines.
2. Describe two methods of obtaining cells for primary cell culture.
3. Mention 2 applications of using cell cultures.
4. Mention 2 limitations of using cell cultures.