technique has been evolved long back, it is only since the dose.
dose. Finally tissue culture is much more amenable to aseptic
Animal Cell Culture Techniques
Cell culture system involves the cells of particular
organ of animal that has been grown in artificial media. Cell
culture system is useful in identification of such
microorganisms which do not grow on artificial media and
require live cells to grow such as viruses. Cell culture
techniques are primarily developed for the viruses for their
further study and development of diagnostics and vaccines.
The cell cultures are primary or continuous cell lines,
that grow under laboratory conditions with proper media and
growth supplements in order to study the virus propagation
and for several other purposes including in vitro toxicological
studies.
Viruses replicate only within the living cells. With the
advent of cell culture technique, it has become easier to grow
the viruses under in vitro condition (cell culture). Viruses are
usually propagated in cell culture, embryonated hen’s egg,
laboratory animals or natural hosts. Although the cell culture
advent of antibiotics that cell culture became a matter of
simple routine technique. Aseptic precautions are still
essential, but the problems of contamination with bacteria,
mycoplasma, fungi and yeasts can be checked by antibiotics
and anti-fungal agents. The term in vitro literally means ‘in
glass’ although today most cell culture is performed in or on
plastic.
Cell culture requires less space, is less expensive and
is more convenient than the use of animals or eggs, although
eggs are still the method of choice in the large scale
preparation of certain vaccines. It is indispensable for the
primary isolation of virus, infectivity assays, biochemical
studies and production of vaccine. An additional advantage of
cell culture systems is that they are completely free from
certain factors which restricts virus multiplication in the intact
animal i.e. virus specific antibodies and non-specific inhibitors
of several kinds. Reproducible infection of a particular kind of
cell by a given virus is thereby facilitated whereas animal may
show various responses to challenge with a standard virus
technique than those involving animals and eggs.
Types of cell culture
Many types of cells undergo only a few divisions in
vitro before dying out, whereas others will survive for up to a
hundred cell generations and some can be propagated
indefinitely. These differences, the nature of which are not
fully understood, give us four main types of cultured cells.
a) Primary cell culture: When cultures are established
initially from tissue taken directly from animals, they
contain several cell types, most of which are capable of
only 5-10 divisions. Due to high cost, inconvenience of
getting fresh tissue each time and variation from batch to
batch, it is not suitable for use in routine diagnostic work
or vaccine production. Furthermore, the donor animals
may harbour the latent viruses which can confuse
diagnosis or contaminate vaccines. But the primary
cultures are very sensitive to many human and veterinary
viruses and still used for primary isolation of these
viruses.
b) Secondary cell culture: When primary cells are again
passaged in vitro, it is called secondary cell culture.
c) Diploid cell strains: These are cells that are capable of
undergoing a number of divisions in culture that is
routinely related to the life span of the species of
animals- about 50 for fetal human cells and about 10 for
fetal cells from horses and cows. Diploid cell strains of
fibroblasts established from human fetuses or embryos
are widely used in human diagnostic virology and vaccine
production, but diploid strains have not been much used
in veterinary vaccine production.

d) Continuous cell line: These are cells of a single type that
are capable of indefinite propagation in vitro. Such
immortal cell lines originate from cancers or by
spontaneous transformation of a diploid cell strain. They
do not bear the close resemblance to their cell of origin as
they undergo many mutations during their prolonged
culture. The cells usually have lost the specialized
morphology and biochemical abilities. Cells of continuous
cell lines are often aneuploid in chromosome number,
especially if of malignant origin. Continuous cell lines
derived from monkey (MA 104, Vero), dog (MDCK), cattle
(MDBK), pig (PK15), Cat (CRFK), mouse (L929, 3T3,
hamster (BHK21), rabbit (RK-13), and others are widely
used in experimental and diagnostic virology. The great
advantage of continuous cell lines over primary cell
cultures is that they can be propagated indefinitely by
subculturing the cells at regular intervals. Like other cells,
they can be preserved for many years when frozen in
serum containing medium with added glycerol or DMSO
and stored at –80°C or –196°C (liquid nitrogen).
Programme
There will be lectures on following topics by Eminent
Scientists during the training programme.
1. Introduction and Design of Animal Cell Culture
2. Preparation of Animal Cell Culture
3. Primary Cell Culture: Principles and Methodology
4. Cell Lines: Sub Culture and Maintenance
5. Cell Culture Related Techniques
6. Applications of Animal Cell Culture Technology
Besides one lecture each day, there will be practicals on
following topics:
1. Preparation of glassware and sterilization
2. Preparation of growth media, buffers etc.
3. Primary cell culture
4. Continuos cell lines
5. Detection of Cytopathology
Training course fee: Rs. 2000/-.
Registration: Send your fee in the form of DD/ Local Bank’s
Cheque in favour of Comptroller, GBPUA&T, Pantnagar along
with details in the following proforma:
1. Name: ……………………………………………………
2. Address:…………………………………………………
3. Phone & Email:…………………………………………
4. Qualification::……………………………………………
5. Experience, if any:………………………………………
6. Organization: ……………………………………………
(Note: Training course fee includes registration kit, lodging and
manual on rabies diagnosis. However, food charges will be paid
as per actual and will be borne by trainees. Registration will be
done on first come first serve basis for only 10 seats.)
Hand’s on Training on Animal
About The Institute Cell Culture Techniques
The Institute of Biotechnology (erstwhile State
Vaccine Institute established in 1903) at Patwadangar (located
on Delhi-Nainital highway 9 km before Nainital in the State of January 27-31, 2010
Uttarakhand in India at an average altitude of 1600 meter). The
small pox vaccine produced and marketed by the erstwhile SVI,
Patwadangar significantly contributed in eradication of small
pox from India. In the year 1957, production of anti-rabies
nerve tissue vaccine of sheep origin was started at the
Institute. Later, the Institute also started production of tetanus
toxoid, which continued till 2004. The Institute stopped
manufacturing of these vaccines due to obsolesce of
technology and non compliance of cGMP norms.
How to reach
Patwadangar is situated 9 km before the Nainital on
Kathgodam – Nainital Highway (NH 87); from Highway
(Baldiakhan bus stop), link road starts to Patwadangar which is
situated at a distance of about 3 km. Kathgodam is well
connected from Delhi, Kolkata, Dehradun through rail and
road. The nearest airport is at Pantnagar which is about 50 km
from Patwadangar. One can reach through rail at Kathogodam
or by air at Pantnagar Airport and then onward journey can be
performed through bus/taxi. Distances to major places:
Nainital (12 km), Kathgodam (29 km), Haldwani (34 km),
Lalkuan (49 km), Delhi (290 km), and Dehradun (335 km).
Climate
Patwadangar is located in Kumaon hills at a height of
1600 meters above sea level. Generally, climate remains cold
and temperature never goes above 300 C in summer. While
visiting this place, woolen clothes are often required. In winter,
sometimes one can also see snowfall.
Contact us
Prof. R.S. Chauhan,
MVSc, PhD FNAVS, FSIIP, FIAVP, Diplomat ICVP
Campus Director
Institute of Biotechnology
(G.B. Pant University of Agri. & Tech.) Institute of Biotechnology
Patwadangar-263128, Nainital
Uttarakhand
G.B. Pant University of Agri. & Tech.)
(
( : 05942- 241129, 241133, 7 : 05942-241231 Patwadangar -263 128 (Nainital)
: : dir_ibtpatwadangar@rediffmail.com,
rs_chauhan123@rediffmail.com
Uttarakhand, India