Why do we need Cell culture? To overcome problems in studying cellular behavior such as: confounding effects of the surrounding tissues. variations that might arise in animals under experimental stress. Reduce animal use. Commercial or large-scale production. Production of cell material: vaccine, antibody, hormone.

1. LECTURE OF SUBJECT :
Dr. sharafaldin Al-musawi
College of Biotecholgy
LECTURE: 2
SUBJECT: Animal Tissue culture
LEVEL: 4

2. Why do we need Cell culture?
Research
To overcome problems in studying cellular behavior such as:
• confounding effects of the surrounding tissues.
• variations that might arise in animals under experimental
stress.
Reduce animal use.
Commercial or large-scale production.
Production of cell material: vaccine, antibody, hormone.

3. Initiation of culture
Tissue
Primary culture
Cell line Continuous cell line
Subculture
Stored Stored
Animal Plant
Finite numbers Indefinite numbers

4. Types of Cell culture
1. Primary Cultures
Derived directly from excised tissue
and cultured either as:
 Outgrowth of excised tissue in culture
 Dissociation into single cells (by
enzymatic digestion or mechanical
dispersion).
Primary
Culture
Preparation

5. Characteristics of Primary Cultures
5
Primary
Culture
Preparation
 Characteristics:
 Morphologically similar to the parent tissue.
 Limited number of cell divisions.
 Best experimental models for in vivo
situations.

6. Advantages & Disadvantages
• Advantages:
• usually retain many of the differentiated characteristics of the cell in vivo
• Disadvantages:
• Initially heterogeneous but later become dominated by fibroblasts.
• The preparation of primary cultures is labor intensive.
• Can be maintained in vitro only for a limited period of time.
• Difficult to obtain.
• Relatively short life span in culture.
• Very susceptible to contamination.
• May not fully act like tissue due to complexity of media.

7. Types of Cell culture
2. Continuous Cultures
 derived from subculture (or passage, or transfer) of primary culture
 Subculture : The process of dispersion and re-culture the cells after they have increased to occupy all of
the available substrate in the culture.
 usually comprised of a single cell type.
 can be serially propagated in culture for several passages.
 There are two types of continuous cultures
 Cell lines
 Continuous cell lines

8. Types of continuous culture
1) Cell lines
• Cell lines derived from primary cultures have a limited life span.
• After the first subculture, the primary culture becomes cell line.
• finite life, senescence after approximately thirty cycles of division.
• usually diploid and maintain some degree of differentiation.
• It is essential to establish a system of Master and Working banks in order to
maintain such lines for long periods.

9. Types of continuous culture
2) Continuous cell lines
 can be propagated indefinitely
 generally have this ability because they have been transformed by:
 tumor cells.
 viral oncogenes
 chemical treatments
 Spontaneously
 disadvantage: having very little of the original in vivo characteristics

10. Transformation VS Transfection
• Transformation
• Spontaneous or induced permanent phenotypic changes resulting from change in DNA
and gene expression that result and effect in:
• growth rate
• mode of growth (loss of contact inhibition)
• specialized product formation
• longevity
• loss of need for adhesion
• Transfection
• Introduction of DNA into a cell (like viral DNA)

11. Cell Culture Morphology
• Morphologically cell cultures take one of two forms:
1. growing in suspension (as single cells or small free-floating clumps) such as: cell lines
derived from blood (leukemia, lymphoma).
2. growing as a monolayer that is attached to the tissue culture flask. Such as: Cells
from solid tissue (lungs, kidney, breast), endothelial, epithelial, neuronal, fibroblasts.
Hela-Epithelial
MRC5-Fibroblast SHSY5Y-Neuronal
BAE1-Endothelial MCF-7 breast
HT1080- kidney 3LL - lungs

12. • Excellent model systems for studying:
 The normal physiology and biochemistry of cells.
 The effects of drugs and toxic compounds on the cells.
 Mutagenesis and carcinogenesis.
• Used in drug screening and development
• Large scale manufacturing of biological compounds
(vaccines, insulin, interferon, other therapeutic protein)
Cell culture application