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ISLAMIC UNIVERSITY –GAZA

Faculty of science
Department of Biotechnology
2nd semester, 2008/2009
Course Syllabus

Course Title Tissue Culture Techniques

Course code
Lecture time
Lab time
Teacher Saib, H al - Owini

1- Course description:
This module is a basic requirement for the department. It provides insights into
the application of animal and plant tissue culture. .It seeks to familiarize students
to the basic principles of tissue culture, and to expose them to its many
applications, the theory production of transgenic plants will be discussed. The
course involves correlation with practical course to ensure full understanding of
the techniques, the lab will provide hands experience in many of the methods
discussed.

2- Course objectives:
- Describe the Equipments used in animal and plant tissue culture.
- Understand the safety procedures need for tissue culture.
- Understand techniques used in tissue culture.
3- Books:
- Culture of Animal Cells, A manual of basic technique, 5th Edi on by Freshney, RI.
WIELY-LISS,2004( optional)

- Experiments in plant ssue culture, 3ed edi on by J.H.Dodds and L.W.Roberts.

Cambridge university press,1999,( optional)

4- Topics to be considered:
A- animal Tissue culture part :
(1) Animal Cell Culture: Historical Background

(2) Importance of and progress in Animal Cell Culture Technology

(3) Biology of animal Cell; basic concepts (Cellular Interactions)

(4) Culture media of animal cells: Serum and Serum Free Media

(5) Cell metabolism and its control in culture media

(6) Mechanism of cell prolifera on and cell death in vitro

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(6) Culturing and Sub-Culturing of Animal Cells

(7) Monitoring and control of cell culture

(8) Cell Differentiation & Cell Movement

(9) Cloning of Animal Cells

(10) Cell Line Preserva on

(11) Cell Line Characteriza on

(12) Mycoplasma: Detection and Control

(13) Monoclonal Antibody Production

(14) product purifica on processes

B- Plant tissue culture :

1- cell theory/totipotency

2- Tissue Culture Methodology

3- Sterile technique

4- Media components

5- Environmental effects

6- Genetic control of culture ability

7- Cell Growth, Mutation and Differentiation Processes in Plant Cultures

8- Cell cycle, population dynamics, growth patterns

9- Differentiation, organogenesis/somatic embryogenesis

10- Undifferentiated (callus, suspension)

11- Organ, meristem, shoot

12- Embryo rescue

13- Somatic embryogenesis

14- Agrobacterium cocultivation

5- Allocation of Marks

30 % Midterm examination, 50% Final examina on and 20 % (Reports, research

projects, Quizzes and Home works).
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PRACTICAL PART OF Tissue Culture Techniques

A- Practical part of animal Tissue Culture Techniques

1- Introduction to Cell culture Lab

2 Aseptic Conditions

3 Equipment, Techniques and Safety in the lab

4 Counting cells with a Hemocytometer

5 Primary fibroblast culture

6 Secondary fibroblast culture

7 Dissecting Mouse and Primary Culture

8 Secondary Culture of mouse cells

9 Primary Cell Culture (Chicken Embryo)

10 Secondary Cell Culture (Chicken Embryo)

11 Tertiary Cell Culture (Chicken Embryo

12 Culture of Human cells (Monocytes) 1

13 DNA ESTIMATION BY HOECHST 33258

14 PROTEIN ESTIMATION BY THE BRADFORD METHOD

15 Culture of Specific Cell Types:

- PANCREATIC EPITHELIUM

16 -KIDNEY EPITHELIUM
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B- Practical part of plant Tissue Culture Techniques

1- Aseptic culture techniques for establishment and maintenance of cultures

2- Preparation of stock solutions of MS basal medium and plant growth regulator stocks.

3- Macro propagation of plants- handout from Practical Botany.

Sow tobacco seeds in sterile chambers ; pour culture tubes according to Nutrition of
callus cultures and culture carrot .
4- Plant nutrition & plant hormones: Effects of hormones on tobacco morphogenesis

5- Propagation from nonmeristematic tissues—organogenesis

6- Plant regeneration-Adventitious organogenesis from tobacco;
continue with making subcultures and/or make media as
needed.
7- Plant genetic transformation-Agrobacterium-mediated
transformation of tobacco in vitro, co-cultivation of leaf
segments; culture maintenance as needed.
8- cryopreservation