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

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 Animal Plant Stored Indefinite numbers
Primary culture
Cell line
Continuous cell line
Subculture
Stored
Animal
Plant
Finite numbers
Indefinite numbers

4. Primary Culture Preparation
Types of Cell culture
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
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
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
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, senesce 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
the disadvantage of having retained very little of the original in vivo characteristics

10. Transformation VS Transfection
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:
growing in suspension (as single cells or small free-floating clumps) such as: cell lines derived from blood (leukemia, lymphoma).
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. Cell culture application
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)