This presentation is about various applications of cell cultures in human/animal viral vaccine production.

1. BT 301 Animal Biotechnology
Applications of cell cultures in
human/animal viral vaccine
production
By,
Aditi Patil
Roll No:11371
MSC-II

2. INTRODUCTION
• Cell Culture is the process by which cells are grown in vitro under defined and
controlled conditions.
• Cell culture can be classified as-
1.Primary Culture.
2.Secondary Culture.
3.Continuous culture.
• Primary cell cultures have limited life span. after certain number of population
doublings cells undergo senescene and stop dividing.
• Cells that divide indefinately called continuous cell lines.
-CHO(Chinese Hamster Ovary)cells
-Vero(Monkey Kidney Cells)
-MDCK(Madine-Darby Canine Kidney Cells)
-PER.C6(Human retinoblast cells)
-MRC-5 (fetal lung cells)

3. Advantages of cell culture
 Vaccines are stable and production is faster as compared to embryo-
-nic chicken eggs.
 Ability to rapidly produce vaccine supplies during impending
pandemic.
 Some strains do not grow well in chicken eggs.
 Avoids egg based allergy reactions.

4. Vaccine
A vaccine is a biological preparation that stimulates production of
antibodies and provides immunity against several diseases.
Vaccine consist of either dead pathogen or live but attenuated
organisms.
 Vaccine induces immunity against pathogen either by antibody
production or by activation of T-lymphocytes.
Consist of : Antigens
Stabilizers
Adjuvants
Antibiotics
Preservatives

5. Types of Vaccines
1. Live Attenuated Vaccine:
- consist of live form of virus that has been artificially weakened
- can revert back to virulent form
- Elicit good immune response, inexpensive.
- Very effective in inducing protection against disease.
- To remain effective,it requires constant refrigeration.
- oral polio vaccine, measles.
2. Inactivated vaccines:
- These made from microorganisms that have been killed by using heat,
radiation or chemicals.
- Reversion to virulent form is high occurrence.
- More expensive to prepare.
- Can be stored without refrigeration.
- Inactivated polio vaccine(IPV)

6. 3.Subunit Vaccine-
-Purified antigen.
-Do not contain live components of vaccine.
-differ from inactivated whole cell vaccine as it contains only antigenic
part of the pathogen.
-e.g.- Haemophillus influenza-B
4.Recombinant Vaccine-
- recombinant vaccines are those in which genes for desired antigen
are inserted into a vector.
- This process induces vector to produce antigen which further get
purified.
-Purified antigen combined with adjuvant results in formation of
effective vaccine.
- e.g. Hepatitis B vaccine
-More boosters are required(immune response is weaker compared to
live,attenuated vaccine)

7. 5.DNA Vaccine-
-Immune response of the body is stimulated by a DNA molecule.
-Induces both humoral and cell-mediated immune response.
-The gene for antigenic determinant of pathogenic organism inserted
into plasmid.
-Genetically engineered plasmid consisting DNA vaccine injected into
host.
-Foreign gene can be expressed from plasmid DNA and it induces
immune response.
6. Toxoid Vaccine-
-Vaccines based on toxin produced by certain bacteria.
-Toxicity is suppresed by heat or chemical treatment.
-No possibility of reversion to virulence.
-This are stable,less suceptible to change in temperature etc.

8. Cell Culture Based Vaccine
Production

9. Applications

10. Vero Cell line
1.One of the most common mammalian continuous cell line used in
research.
2. Anchorage-dependent cell line.
3. Licensed in US for production of live(rotavirus,smallpox) and I
inactivated(poliovirus) viral vaccine.
4.Also used for production of other viruses such as Rabies
virus,reovirus.
Fluroscent image of Vero Cells

11. Japanese Encephalitis-
1. JEV is a flavivirus related to dengue,yellow fever spread by
mosquitoes.
2. Inactivated Vero cell culture derived vaccine (Ixiaro) was developed.
3. Vaccine is based on attenuated strain of JEV which grow on Vero cell.
4. Purified product adjuvented with 0.1% aluminium hydroxide.

12. Rabies
1. Rabies virus causes inflammation of brain.
2. Symptoms such as violent movements.uncontrolled exitement,
confusion, loss of conciousness finally death.
3. Vaccine is produced using vero cells adhered to microcarriers
and cultivated in biorector in serum free medium.
4. Vaccine purification by chromatography and inactivated using
beta-propiolactone.
1. IMOVAX vaccine prepared using human diploid cells
(MRC-5 strain)
2. Concentrated by ultrafiltration.
3. Inactivated by beta propiolactone.

13. Polio-
1.Inactivated polio virus vaccine prepared using sabin strain.
2. Cell culture used: PER-C6.
Production of sabin virus in PER-C6 Cells
Harvesting(removal of cell debris),filtration(to
remove impurities)
Virus purification by size exclusion chromatography
Inactivation at 37°C in glycine,foraldehyde.

14. Livestock Diseases

15. Pseudorabies-
1. Pseudorabies virus(PRV) is double stranded DNA-based swine
virus.
2. Pseudorabies also called as “Aujeszky’s disease”
3. Caused by Suid herpesvirus 1(SuHV1).
4. Viral disease of pigs.also affects dogs, cattle, goats etc.
5. Inactivated and attenuated vaccines are available.
6. Swine testis cells are use for vaccine production.
7. Microcarriers as well as suspension culture is used for
vaccine production.

16. Foot and Mouth disease-
1. FMD is highly contagious disease affects cattle and pigs.
2. 1st FMD vaccine was developed (1938) based on formaldehyde
inactivated virus harvested from artificially infected cattle.
3. Mowat-Chapman (1962) found FMD virus multiply efficiently in baby
hamster kidney (BHK) cells.
4. Suspension/ adherent cell culture system can be used.
BHK cell culture
Virus inoculation
Inactivation using Aziridine compounds (ethyleneimine)

17. Harvest and concentration.
Formulation with adjuvant(Saponin,aluminium hydroxide gel/oil
emulsion)

18. References-
1. Craig R. Barett et al. The BRIDGE,National Academy of
Engineering,Washington.
2. Recommendation for evaluation of animal culture as substrates for
manufacture of biological medicinal products.
3. Viki Bockstal et al. 2018,An inactivated poliovirus vaccine using sabin
strains produced on serum-free PER-C6 cell culture platform is
imuunogenic and safe in non-human primate model.
4. P Noel Barrett et al. 2009, Vero cll platform in vaccine production:
moving towards cell-culture based viral vaccines,Vaccines8(5)
607-618.
5. J.Luborth et al. 2007,Veternary Vaccines,26(1),179-201.
6. C.M. Freuling,2017 Vaccines against pseudorabies virus(PrV)