The document summarizes the nutritional requirements of cultured cells. It discusses that cells need nutrients like amino acids, vitamins, inorganic ions, trace elements, monosaccharides, and growth factors to survive and grow in culture. Specific requirements are outlined, including essential amino acids, glutamine, salts like sodium, potassium and calcium, and glucose as an energy source. Serum provides additional growth factors, minerals, lipids and hormones to support cell growth. Characteristics of cell culture and applications are also briefly mentioned.

1. Biotechnology Skill Enhancement Program (BiSEP)
Domain: Fermentation and Bio-processing
Seminar Presentation
Seminar Topic: Nutritional Requirements of Cultured cells
Name: SACHIN .B .H
Date: 13/11/2019
Department of Biotechnology, GUK

2. Synopsis
1) Introduction
2) Animal cell culture
3) Cell culture media
4) Nutrional requirements of cultured cells
5) Characterstics

3. INTRODUCTION
Cell culture can be defined as the process of
cultivating cells and tissue outside the body of an
organism(invitro) in an artificial environment, which
stimulates the invivo conditions such as temparature,
nutrition and from microorganisms.
Cell culture is the process by which cells are grown
under controlled conditions, generally outside of
theire natural environment.
Cell culture was first successfully undertaken by Ross
Harrison in 1907.

4. ANIMAL CELL CULTURE
1) In vitro cultivation of animal organs, cells,and
tissues at defined temperature using an incubator and
supplemented with a medium containing cell nutrients
and growth factors is collectively known as Animal cell
culture.
2) And will continue to grow if supplied with
appropriate conditions and nutients.
3) Culture process allows the single process to act as
an independent unit.
4) Cells are capable of dividing.

5. Terminologies
1) Primary Cell culture: When cells are surgically
removed from an organisms and placed into a suitable
culture environment they will attach, divide and grow
2) Cell Line: When the primary culture is subcultured
and they show an ability to continuously propagate.
3) Anchorage depedency: Cells grow as monolayers
adhering to the substrate (glass/plastic).
4) Passaging/subculturing: The process of spliting the
cells.
5) Finite cells: When the cells has finite life span.

6. Cell Culture Media
Cell culture media generally comprise an appropriate
source of energy and compounds which regulate the
cell cycle.
A typically culture medium is composed
Amino acids
Vitamins
Inorganic salts
Glucose and serum

7. Nutritional Requirements of
cultured cells
Nutrients:
Components in food that a cell uses to survive and grow are
known as nutrients.
Nutritional Requirements:
1) Amino acids ( arginine, leucine etc)
2) Vitamins ( Riboflavin, biotin, vitamin B12)
3) inorganic ion ( sodium, potassium, calcium)
4) Trace elements ( molybdenum, copper)
5) Monosaccharide ( Sugar, glucose, galactose)

8. Serum
Serum contains
Growth factors
Minerals
Lipids
Hormones
Many of which may be bound to protein
The sera used most in tissue culture are bovine
calf, fetal bovine, adult horse, and human serum
Calf (CS) and fetal bovine (FBS) serum bovine are the
most widely used, the later particularly for more
demanding cell lines and for cloning.

9. Protein
1) Protein are Required as carriers of minerals, fatty
acids, and hormones.
2) Albumin as a carriers of lipids, minerals, and
globulins.
3) Fibronectin (cold-insoluble globulin), which
promotes cell attachment.
4) Fetuin in fetal serum enhances cell attachment.

10. Growth factors
1) Natural clot serum stimulates cell proliferation
more than serum from which the cells have been
removed physically
2) This increased stimulation due to growth factors.
3) PDGF- Platelet derived growth factor, from the
platelets during clotting.
4) PDGF Stimulates growth in fibroblasts and glia.

11. Amino acids
The essentials amino acids are required by Cultured cells
1) plus cystein
2) Arginine
3) Glutamine and tyrosine
4) The concentration of amino acids usually limits the
maximum cell concentration attainable, and the balance
may influence cell survival and growth rate.

12. Glutamine
1) Glutamine is required by most cells, although some
cell lines will utilize glutamine.
2) Glutamine is also used in cultured cells as a source
of energy and carbon.
3) Glutamine is unstable in culture medium with a
half-life of between 3 and 5 days, and it generates
ammonia, which can be toxic as a by product.

13. Salts
1) The salts are chiefly those of a Na+, K+, Ca2+, and
HCO3-and are the major components contributing to
the osmolality of the medium.
2) Ca2+, are required by some cell adhesion
molecules, such as the cadherins.
3) Calcium is reduced in suspension cultures in order
to minimize cell aggregation and attachment.
4) Na+, K+, and Cl- regulate membrane potential.

14. Glucose
1) Glucose is included in most media as a source of
energy.
It is metabolized principally by glycolysis to
form pyruvate, which may be converted to lactate or
acetoacetate and may enter citric acid cycle and be
oxidized to form CO2 and water.

15. Trace elements
1) Trace elements are often supplemented to serum-
free media to replace those normally found in serum.
2) Trace elements like copper, zinc, selenium, are
needed in minute amounts for proper cell growth.
3) These micronutrients are essential for many
biological process, e.g. the maintenance of the
functionality of enzymes.

16. Contd…
Animal cell culture media vary in their
complexity but most contain:
1) Amino acids : 0.1-0.2mM
2) Vitamins : ca.1µM
3) Salts : Nacl 150mM
Kcl 4-6mM
CaCl 1mM
4) Glucose : 5-10mM

17. CHARACTERISTICS
1) Animal cells can grow in simple glass or plastic
containers in nutrive media but they grow only to
limited generation.
2) The maintenance of growth of cells under
laboratory conditions in suitable culture medium is
known as PRIMARY CELL CULTURE.
3) Cells can grow as adherent cells ( anchorage
dependent must be cultured while attached to a solid
or semi-solid substrate.
4) or as suspension culture ( anchorage independent
can be grown floating in the culture medium)

18. Application of cell culture
1) Excellent model systems for studying:
- The normal physiology, cell biology and biochemistry of cells.
- The effects of drugs, radiation and toxic compounds on the cells
- Study mutagenesis and carcinogenesis.
2) Used for gene transfer studies:
- Large scale manufacturing of biological compounds .
- (vaccines, insulin, interferon, other therapeutic protien)

19. REFERENCES
1) Butler's. (2005) Animal cell culture: recent
achievements and perspective in the
production of biopharmaceuticals.
2)Freshney R.I (2010) Culture of animal cells
3) Gngal.S. (2007). Principle and practice of
animal cell culture.