Cell culture based vaccine?? Cell cultures involve growing cells in a culture dish, often with a supportive growth medium. A primary cell culture consists of cells taken directly from living tissue, and may contain multiple types of cells such as fibroblasts, epithelial, and endothelial cells. In the United States, 10 different vaccines for chicken pox, hepatitis A, polio, rabies, and rubella are cultured on aborted tissue from two fetal cell lines known as WI-38 and MRC-5. These vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio, rabies, rubella, measles/rubella, mumps/rubella, and MMR II (measles/mumps/rubella).

1. CELL CULTURE BASED VACCINE
By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )

2. Introduction
 Cell culture based vaccine??
 Cell cultures involve growing cells in a culture dish, often with a
supportive growth medium. A primary cell culture consists of cells
taken directly from living tissue, and may contain multiple types
of cells such as fibroblasts, epithelial, and endothelial cells.
 In the United States, 10 different vaccines for chicken pox,
hepatitis A, polio, rabies, and rubella are cultured on aborted
tissue from two fetal cell lines known as WI-38 and MRC-5. These
vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio,
rabies, rubella, measles/rubella, mumps/rubella, and MMR II
(measles/mumps/rubella).

3. History
 PCCs of monkey kidney cells have been used for the production of
inactivated and oral poliomyelitis vaccines since the 1950s.
 In 1954, an experimental adenovirus vaccine was being developed,
and human tumour cells (HeLa) were rejected as the cell substrate in
favour of ”normal” cells.
 The first requirements for cell substrates were published by WHO in
1959 for the production of inactivated poliomyelitis vaccine in PCCs
derived from the kidneys of clinically healthy monkeys.
 In the 1960s, human diploid cells (HDCs) were developed and
proposed as an alternative to primary monkey kidney cell cultures for
polio virus vaccine production as well as for other viral vaccines.
 WHO Requirements for Continuous Cell Lines used for Biologicals
Production were published in 1987.
 During the 1990s, and on into the 2000s, a variety of CCLs were
explored as cell substrates for biological products.

4. Types of animal cell substrates
1. Primary Cell Cultures (PCCs)
o Major successes in the control of viral diseases, such as poliomyelitis,
measles, mumps and rubella, were made possible through the wide
use of vaccines prepared in PCCs, including those from chicken
embryos and the kidneys of monkeys, dogs, rabbits and hamsters.
Advantages:
(a) Easy to prepare using simple media and bovine serum;
(b) Broad sensitivity to various viruses, some of which are
cytopathic.
Disadvantages:
(a) Contamination by infectious agents is a higher risk than
with DCLs and CCLs;
(b) They cannot be tested as extensively as DCLs or CCLs.

5. Types of animal cell substrates
2. Diploid Cell Lines (DCLs)
The practicality of using human DCLs for the production of viral vaccines was
demonstrated in the 1960s. DCLs of human (e.g., WI-38, MRC-5) and
monkey ( i.e., FRhL2 ) origin.
Advantages
(a) They can be well characterized and standardized;
(c) Unlike the CCLs and SCLs , DCLs are not tumourigenic and therefore do not
raise the potential safety issues associated with CCLs and SCLs.
Disadvantage
(a) In general, they have more fastidious nutritional requirements than other cell
substrates;
(b) They may be difficult to adapt to serum-free growth;
(c) They are more difficult than CCLs to transfect and engineer, and require
immortalization before they can be engineered;

6. Types of animal cell substrates
3.Continuous cell lines (CCLs)
CCLs have the potential for an apparently indefinite in vitro life
span and have been derived by the following methods:
a) Serial subcultivation of a PCC of a human or animal tumour (e.g.,
HeLa cells);
(b) Transformation of a normal cell having a finite life span with an
oncogenic virus or viral sequence (e.g., B lymphocytes
transformed by EBV or transfected with viral sequences such as in
PER.C6);
(c) Serial subcultivation of a primary-cell population derived from
normal tissue that generates a dominant cell population having an
apparently indefinite life span, often described as spontaneous
transformation (e.g., Vero, BHK-21, CHO, MDCK, Hi5);
(d) Fusion between a myeloma cell and an antibody-producing B
lymphocyte to produce a hybridoma cell line.

7. Advantages:
(a) Characterized extensively and their culture conditions
standardized;
(b) Grow more easily than DCLs using standard media,
(d) Most can be adapted to grow in serum-free medium;
(e) Can be grown on micro-carriers for large-scale production
in bioreactors;
(f) some can be adapted to grow in suspension cultures for
large-scale production in bioreactors.
Disadvantages
(a) CCLs may express endogenous viruses, and some are
tumourigenic in immuno-suppressed animal models;
(b) Theoretical risks identified by the 1986 Study Group (e.g.,
nucleic acids, transforming proteins, and viruses) need to be
taken into account.

8. GENERAL STEPS OF VACCINE PRODUCTION FROM ANIMAL
CELLS

9. VACCINES FROM CELL CULTURE
 MMR Vaccine
 Polio vaccine
 Chickenpox (Varicella) vaccine
 Rabies vaccine
 Recombinant protein vaccines
 Influenza vaccine
 Dendritic Cell-Based Vaccination in Solid
Cancer

10. MMR Vaccine
Disease Immunized Virus Strain Propagation cell Growth Medium
Measles
Enders' attenuated
Edmonston strain
chick embryo cell
culture
Medium 199
Mumps
Jeryl Lynn(B level)
strain
chick embryo cell
culture
Medium 199
Rubella
Wistar RA 27/3 strain
of live attenuated
rubella virus
WI-38 human diploid
lung fibroblasts
MEM (solution
containing buffered
salts, fetal bovine
serum, human serum
albumin and
neomycin, etc.)

11. Polio , Chickenpox and Rabies vaccine
Disease Immunized Virus Strain Propagation cell Growth Medium
Poliomyelitis or
polio
Type 1 (Mahoney),
Type 2 (MEF-1),
Type 3 (Saukett)
vero cells, a
continuous line of
monkey kidney
cells
Eagle MEM
modified medium,
M-199
Chickenpox Oka/Merck strain
(varicella virus)
Human diploid
cell cultures
(WI-38, MRC-5)
MEM
Rabies
Attenuated
Pitman-Moore
L503 strain
Attenuated Wistar
strain
Human diploid
cell
chicken embryo
Vero cell
Eagle MEM
modified medium,
M-199

12. iNfluenza vaccine

18. (VLPS)-based Influenza Vaccines

21. DC-based cancer vaccine
• Dendritic cells (DC), a leukocyte population,
represent unique antigen-producing cells capable
of sensitizing T cells to both new and recall
antigens.
• DCs are the most potent antigen-presenting cells
(APS).
• Tumor antigens in different forms (DNA, RNA,
proteins, peptides, viruses, cell lysates) become
immunogenic when presented to T-lymphocytes by
DCs.
• Immunization with ex vivo generated DC has proven
feasible and permits the enhancement as well as
the dampening of antigen-specific immune
responses in man.
• Several DC-based clinical trials have demonstrated

23. DC loading and activation
 Ex vivo generated dendritic cells(DC) can be loaded
with antigens and re-infuse to the patients, or they
can be used for ex vivo expansion of anti-tumor
lymphocytes.

24. Personalized Cancer Vaccine
More than 150 DC-based clinical studies for the
treatment of solid or hematological malignancies
have been reported in 2008 including;
 Melanoma (〉 40 published clinical studies)
 Prostrate cancer (20)
 Renal cell carcinoma (16)
 Breast cancer (12)
 Multiple myeloma (9)
 Leukemia (9)
 Colorectal cancer (9)
 Glioma (9) Nencioni, A. et.al. Critical Reviews
in Oncology/Hematology
65(2008), 191-199

25. Advantages
 Eliminate the need for embryonated chicken eggs
from managed, biosecure flocks.
 Combine and automate upstream and downstream
processes.
 Reduce the potential for contamination by viable and
nonviable particulates.
 Eliminate the four- to six-month lead times for the
organization of egg supplies.
 They have faster, high-volume start-up times for
production.
 Higher initial purity.
 They could supplement seasonal vaccine supplies
when multiple strain changes are necessary.
 They would substantially increase global stockpiles of
pandemic influenza vaccines.

26. Potential risks
 Viruses and other transmissible agents
 Cellular DNA
 Cellular RNA
 Limitations of Vaccine Production using cell
culture
 Not all infectious agents can be grown in culture
 Animal/human cell culture is expensive
 Yield of viruses from cultures can be low
 Safety precautions for culture of live agents

27. The top biotech companies in India
 Biocon : Bangalore, Karnataka
 Serum Institute of India :Pune, India
 Panacea Biotech : Mumbai
 Shantha Biotech : Hyderabad
 Indian Immunologicals : Hyderabad, Ooty and Rajkot
 Bharat biotech : Hyderabad
 Syngene International : Bangalore

28. REFERENCES
 Springer.com
 WHO official website
 www.biopharminternational.com