Download Strictly Use the provided Space only!!! If you do not place your

Lecturer: Dr. Raida Khalil
Faculty of Science
Internal Examiner :
Department of Biotechnology & Genetic
Engineering
Student Name:
Student Number:
Course Name: Animal Tissue Culture
First Semester: 2010/2011
Course Number: 240472
Section: 1
Date: 18th Jan 2011
Final Exam
Time: 14:00 pm
Duration: 100 minutes
Number of Pages:
6
Number of Questions: 10
Mark:
/50
Strictly Use the provided Space only!!!
If you do not place your answers in the appropriate locations, I may not find them!!!
"The main object of education is to keep continually asking questions"
Q1. Suppose your desired cell number is 104 and you have 5 ml of your cell suspension
containing 2.3 x 10^6 cells/ml, find out how much medium you need to get the desired
cells? (2 pts)
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All the best, Dr. Raida W. Khalil
Q2. Match the terms in column A with proper definition in Column B:(8 pts)
Column A
Column B
Aseptic
a- where all the cells are in contact all around their
periphery with other cells, and no available substrate is left
uncovered
Passage
b-Free of Microbial infection
Fibroblast
c-a culture that has been propagated by subculture but is
only capable of a limited number of cell generation in vitro
before dying out
Laminar flow
d- the transfer or subculture of cells from one culture vessel
to another.
cell line
e- The flow of fluid that closely follows the shape of a
streamlined surface without turbulence
Immortalization
f- a proliferating precursor cell of the mature differentiation
fibroblast
Confluent
g- an increase in effect produced by a given stimulus
Induction
h- the acquisition of an infinite life span
Q3. Order the elements of selection in the evolution of Cell lines, using Arabic
Number(5pts)
Element
Order
Propagation as cell line
First subculture
Isolation
Primary Culture
Transformation
2
All the best, Dr. Raida W. Khalil
Q4.The following items used in the tissue culture and should be stored at particular
temperature, indicate the OC for each( 5pts)
Item
OC
PBS
RPMI and
DMEM media
Pipettes
Fetal calf serum
Culture flasks
Q5. This figure represents-------------------------------? (2 pts)
Q6. Preventing cell culture contamination has long been the dream of many
researchers! What are the consequences of cell culture contamination?( 3 pts)
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All the best, Dr. Raida W. Khalil
Q7.T here two major sources of cell culture contamination, Biological and Chemical.
A- What are the sources of Biological contaminants? (2pts)
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2-------------------------------------------------------------------------------------------B- Biological contaminants subdivided into two groups based on the difficulty of
detection them in culture? (2 pts)
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2------------------------------------------------------------------------------------------------C- How can cell culture contamination be controlled? (3 pts)
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3-------------------------------------------------------------------------------------------Q8: A mouse cell lacking the enzyme thymidine kinase was fused with primary human cell
containing this enzyme. The cells then were grown in the absence of thymidine. After several
generations of growth, several cell clones were isolated and assayed for thymidine kinase. In
addition, the human chromosomes stained by these hybrids were identified based on their
banding patterns.
1-Based on the resulting data shown in Table 1, what can you conclude about the
chromosomal location of the gene encoding thymidine kinase? (2pts)
-----------------------------------------------------------------------------------------------2-How could you confirm your conclusion experimentally? (2pts)
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All the best, Dr. Raida W. Khalil
Table 1
Clone
Thymidine Human Chromosome
Number
Kinase*
1
+
9,11,17.20
2
+
3,11,17,19
3
2,3,11,20
4
+
16,17,21
5
+
9,12,17,19
6
9,16,21
7
+
2,17
* (+) indicates presence and (-) indicates absence of thymidine kinase
Q9 You have been asked to analyze chromosomes of the HIT 2 Cell lines grown in flask and
lysine coated.. Although the preparation of metaphases appears to be difficult. you are
confident that it can be done readily. Describe your stepwise approach in brief? (5pts)
Step
A-
Chemicals, equipments ------etc
BC-
D-
E-
Q10. You have learned and practice MTT Assay in week 10 and gained brilliant experience.
Accordingly, Pharmaceutical company request in vitro Cytotoxicity test for four novel
drugs (A, B, C,D) used for Diabetes treatment.
Dear Student design your MTT assay, proposed your own readings (tabulate data), plot
curve and finally make a conclusion! (10 pts)
5
All the best, Dr. Raida W. Khalil
Do not forget the following tips:
*INS I cell lines( pancreatic cell lines) used,* Four drugs diluted in PBS
*Do not forget to use control!,* Analyze Data: do not forget to plot a curve!
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End of paper
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All the best, Dr. Raida W. Khalil