It explains the basics of cell culture

1. Lab 1: Primary cell culture and cell lines
Molecular and cellular pathogenesis/practical
4th stage- 2nd semester
Dr. Amir Hani Raziq
Asst. Prof. In Genetic Engineering and Biotechnology
Kurdistan Regional Government – Iraq
Ministry of Higher Education & Scientific Research
Cihan University-Duhok
College of Health Sciences
Department of MLD

2. What is Cell Culture?
Cell culture is the removal of cells from an animal or plant and their subsequent growth in
a favorable artificial environment.
The cells may be removed from the tissue directly and disaggregated by enzymatic or
mechanical means before cultivation, or they may be derived from a cell line or cell strain
that has already been established.

3. Tissue disaggregation by enzymes
to obtain single cells.
Most commonly used enzymes
are:
• Trypsin.
• Collagenase.
• Dispase.

4. Mechanical disintegration of
tissue to create the primary
explant

5. Cells can be purchased as
frozen vials
Human Hepatocyte Primary
Cell Culture - Frozen Vial
(liquid nitrogen), -169 oC

6. Major developments in cell culture technology
• First development was the use of antibiotics which inhibits the
growth of contaminants.
• Second was the use of trypsin to remove adherent cells to
subculture further from the culture vessel
• Third was the use of chemically defined culture medium.

7. Cell cultures or cell lines can be of two types based
on their adhering ability:
• Suspension cell culture or suspension cell lines.
• Adherent cell cultures or adherent cell lines.
Adhering cells
Adhering and detached cells

8. What is the cell culture used for?
• Studying basic cell biology, interactions between pathogens and cells, effects of
drugs on cells, and nutritional studies.
• Toxicity testing.
• Study the effects of new drugs.
• Cancer research.
• Many other scientific and applied fields.

10. Types of cells
• On the basis of morphology (shape & appearance) or on their
functional characteristics. They are divided into three.
• Epithelial like-attached to a substrate and appears flattened and
polygonal in shape.

11. • Lymphoblast like-cells do not attach remain in suspension with a
spherical shape.

12. • Fibroblast like-cells attached to an substrate appears elongated and
bipolar.

13. Types of cultures
• Primary culture: when cells are isolated
from the tissue and proliferated under the
appropriate conditions until they occupy all of
the available substrate (reach confluence). At
this stage, the cells have to be subcultured
(passaged) by transferring them to a new
vessel with a fresh growth medium to provide
more room for continued growth.

14. Primary culture
• Primary cells have a finite life span.
• Primary culture contains a very heterogeneous population of cells.
• Sub culturing of primary cells leads to the generation of cell lines.

15. Finite and continuous cell lines
• Normal cells divide to a limited number of times then they stop dividing. This
process is called senescence (aging); these cell lines are known as finite.
• Some cell lines become immortal through a process called transformation.
• Transformation can occur spontaneously or can be chemically or virally induced.
• When a finite cell line undergoes transformation and acquires the ability to divide
indefinitely, it becomes a continuous cell line.

16. THANK YOU