The growing interest in products from animal cells has caused an
research effort for the development of media for cell cultivation. The
components in the media used for cultivation of animal cells vary
the character of the cells, and the cultivation method. Basic components
of an energy source, nitrogen source, vitamins, fats, fatty acids and
components, inorganic salts, nucleic acid, antibiotics, oxygen, pH
buffering systems, hormones, growth factors serum, and extensive efforts
directed toward developing serum-free media, protein media, or
chemically defined media (e.g., MEM—minimum essential media).
Almost 50% of the biologicals produced today or planned to be produced
future are of animal cell origin. Therefore, there is an increasing
developing technologies for cultivation of animal cells and production
of a wide
spectrum of biologicals. The worldwide activities and market of
mammalian cells were recently received.
Although the major achievements in the field of animal cell cultivation
been accomplished in the last 3 decades, it has a long history of about
hundred years. Apart from the development of various types and size of
culturing vessels, research and development of optimal media for cell
also carried out among most groups involved in the field of animal cell
cultivation and production of biologicals. Media used for cell
considered to include 2 major parts.
Water for Animal Cell Media
- Essential basal ingredients that fulfill all cellular requirements for nutrients,
known as the basal growth medium.
- A set of supplements that satisfies other types of cellular growth
requirements and makes it possible for the medium to grow.
Out of the most important points for consideration when preparing media
required high quality of the water. Water used for culture media should
pyrogen-free (especially if the product is for human or animal use it
have resistance of 1.5–2.0 m. Ohms, indicating a low salt content.)
It is highly recommended to use fresh ultrapure water, and not store water
since in some storage tanks, organic materials, or irons from plastic or glass
may dissolve in the water.
Purity of Chemicals, Stability, and Shelf Life
Chemicals of the highest purity are required for preparation of media.
chemicals, although pure, inevitably contain traces of contaminants.
traces may be toxic (like Hg). With regard to stability of media
inorganic chemicals are indefinitely stable. Vitamins are the least
Hormones, several antibiotics, and growth factors are recommended to be
(–20°C) or refrigerated (0°C–4°C).
Several ingredients used in animal cell culture media are known for
instability, e.g., ascorbic acid and glutamine. Most factors affect the
shelf life of
among them are the following: Natural decay rates of unstable compounds,
pH, moisture, storage temperature, access of oxygen, and an exposure
near-ultraviolet, day light, or inflorescence light. Most media should
and in a dark place. Storage of media by freezing may cause loss of
some purely soluble ingredients. Powdered media may be stored for
Basic Components in Media
Sera in Animal Cell Media
- Energy sources—Glucose, fructose, amino acids.
- Nitrogen sources—amino acids.
- Vitamins: mainly water-soluble vitamins—B and C.
- Fat and fat-soluble components: fatty acids, cholesterols.
- Inorganic salts: Na+, K+, Ca2+, Mg2+
- Nucleic acid precursors
- pH and buffering systems.
- Hormones and growth factors.
Sera is the most important and most problematic component in animal cell
During more than 3 decays, sera has been an essential medium component
with the following functions:
- Provides nutrients.
- Provides proteins that solubilize essential nutrients that do not dissolve
- Binds essential nutrients that are toxic when present in excessive amounts
and releasing then slowly in a controlled manner.
- Provides hormones and growth factors.
- Modulates the physical and chemical properties of the medium (viscosity,
rate of diffusion)—protect cells in agitated culture.
- Has a pH-buffering function.
Despite these advantages, there are several problems associated with the
use of serum for cell cultures.
- Serum is the most expensive component.
- Being highly viscous, sera slows down the sterilization by filtration of the
- From time to time, there is a shortage in world supply of sera.
- Possible availability of contaminants.
For example, Mycoplamas, viruses.
- Availability of serum in media increases the complicity of the downstream
processing of the desired biological media.