Tissue Culture

(BTEC4331)
Fall 2014

Dr Saeb Aliwaini

Office hours:

Saturday: 12-2 Tuesday: 11-12 Wednesday: 12-2

If necessary, appointments can also be scheduled. I can also be contacted by e-mail.

Text: Culture of Animal Cells: A Manual of Basic Technique (6thEdition) R. Ian

Freshney.

Objectives: This course is designed to provide students with the basic skills needed to
manipulate mammalian cells in culture. Laboratory skills in cell growth, cell passage,
gene transfection, and enzyme assays will be acquired.

Testing: All the material covered in the exams will be based on information from the
textbook, lectures and laboratory exercises. In order to do well in this course, the
assigned reading must be read before lecture to help you grasp the problems and to
ask for clarification of concepts when necessary.

Grades: Midterm Exams 30%

Final Exam 50%
Lab assignments 20%
Total = 100 %

Laboratory Lab: notes and post-lab reports are required for labs on the day they are
due.
Reports:
Outline and rules for lab reports: The following outline should be used for the laboratory
(computation) notebook and for the Post-lab write-ups. You must follow this outline and
rules. For each lab assignment, you will include your handwritten rough notes and
calculations. Automatic 10 points your assignment if handwritten lab notes are not
included in lab reports. If your rough lab notes an calculations are handed in as typed,
you will still lose 10 points.
A. Title of Experiment and Date
B. Purpose of Experiment
C. Materials (include calculations for all solutions)
D. Methods (write pre-lab on the left half of the page and record procedures performed
on right half during experiment.)
E. Results: Record all results in the notebook. All pictures and printouts are raw data
and should be taped into notebook and properly labelled.
F. Conclusions: Record your conclusions for the experiment in the notebook.

Course Schedule:
Topic chapter
1- Orientation /Introduction to Tissue Culture /Aseptic Technique ( 1 and 5 )
2- Biology of Cultured Cells/Equipment (2 ,3and 4)
3- (Counting) Safety /Quantitation (6 and 20)
4- Cryopreservation ( 19)
5- Primary cultures/Subculture & Cell Lines (11 and 12)
6- Specialized Techniques/Molecular Techniques ( 27)
7- Preparation & sterilization (10)
8- Culture Vessels / Defined (7and 8)
9- Cloning and Selection/ Cell Separation (13 and 14)
10- Characterization/Differentiation (15 and 16)
11- Cytotoxicity/ Organotypic Culture ( 21 and 25)
12- Contamination (18)

Labs Lab 1: Sterile Technique and media preparation

Lab 2: Culturing adherent cell lines and photography.
Lab 3: Cell Growth Curve and Drugs treatment
Lab 4: Freeze Downs and thawing, contaminations (Mycoplasma test)
Lab 5: Gene Transfection
Lab 6: Isolating Protein
Lab 7-8: SDS PAGE, Transfer, Western Blot.
Lab 9-10: Immunofluorescence & Microphotography
Lab 11: Cell migration assay
Lab 12: MTT assay
Lab 13: cell death assay
Lab 14: Luciferase assays
Lab15: Flow cytometry, Cell cycle analysis and apoptosis.
Lab 16: Quantitative real-time PCR (qRT-PCR)
Lab 17: Chromatin immunoprecipitation (ChIP) assay