KATHMANDU UNIVERSITY

Marks Scored:
End Semester Examination[C]
2014
Level : B. Tech. Course : BIOT 304
Year : III Semester : I

Exam Roll No.: Time: 30 mins F. M. : 20

Registration No.: Date :

SECTION "A"
[20Q  0.5= 10 marks]

Mark “X” in the most appropriate box.

1) Cells are dissociated from tissues by mechanical means and by enzymatic digestion using
[ ] hydrolyzing enzymes [ ] oxidizing enzymes
[ ] proteolytic enzymes [ ] none of the above

2) For the Haemocytometer slide, the volume of the chamber is,

[ ] 0.001mm3 [ ] 0.01mm3 [ ] 0.1mm3 [ ] 1mm3

3) Which of the following is a not stage of Primary culture?

[ ] Acquisition of the sample [ ] Isolation of the tissue
[ ] Dissection/disaggregation [ ] Subculture

4) Beneficial equipment in cell culture is,

[ ] Balance [ ] -20 freezer
[ ] Fluorescence microscope [ ] Haemocytometer

5) The choice of cell culture vessel depends on:

[ ] Cell yield [ ] necessity of CO2 or not
[ ] Culture is in suspension or grown as a monolayer [ ] All of the above

6) To increase viscosity the substance added to cell culture medium is

[ ] Serum [ ] Carboxy methyl cellulose
[ ] Hanks salt [ ] Earls salt

7) The number of doubling the cell population has undergone is known as…………….
[ ] Passage number [ ] Generation number
[ ] Split ratio [ ] All of the above

8) Who is regarded by some as father of cell culture

[ ] Rous and Jones [ ] Alexis Carrel [ ] Carrel and Baker [ ] Ross Harrison

9) Which type of cell culture is most reproducible

[ ] Organ [ ] Explant [ ] Cell [ ] All of the above

10) Which of the following is most unstable in cell culture medium.

[ ] Glutamine [ ] Threonine [ ] Aspartate [ ] Serine
11) Which of the radioactive nucleotide is used for measurement of DNA synthesis
[ ] 1H thymidin [ ] 2H thymidin [ ] 3H thymidin [ ] Non of the above.

12) _________ jacketed incubators distribute heat evenly around and hold their temperature
longer.
[ ] Air [ ] Water [ ] Steam [ ] Hot air

13) Integrins, mediate cell–matrix adhesion via a specific motif usually containing the
[ ] GDR motif [ ] RGD motif [ ] RDG motif [ ] All of the above

14) __________ in trypsin chelates Ca ions and thus helps dissociation of Ca dependent
cadherins.
[ ] Protease [ ] RNAase [ ] DNAse [ ] EDTA

15) Cell attachment is regained in Glass surface by treating with ____ mM magnesium acetate.
[ ]1 [ ]2 [ ]3 [ ]4

16) The most commonly used cryoprotectant, DMSO, is added at final concentration of ______ .
[ ] 5% [ ] 10% [ ] 15% [ ] 20%

17) Sterility testing of culture medium can be carried out by

[ ] Autoclaving [ ] Membrane filtration
[ ] Incubation [ ] All of the above

18) Cells with a _______ oxygen requirement do better in a shallow medium (2mm)
[ ] Low [ ] No
[ ] High [ ] Non of the above

19) Which of the following is not controlled in Cell culture?

[ ] pH [ ] Relative Humidity
[ ] Volume [ ] Osmotic pressure

20) T25 flask is called so because

[ ] It can hold 25ml of media [ ] It can hold 25x106cells
[ ] Its surface area is 25cm2 [ ] It surface area is 25mm2

SECTION "B"
[10Q  1= 10]
Fill in the blanks

1) α2-macroglobulin present in serum helps to inhibit _______________.

2) ___________________is a yellow water-soluble tetrazolium dye that is reduced by live, but

not dead, cells to a purple formazan product that is insoluble in aqueous solutions.

3) If cloning is performed in __________________ , there is no physical barrier between

colonies and this barrier must be created by removing the medium and placing a stainless steel

or ceramic ring around the colony to be isolated.

4) One way to maintain cell at clonogenic density but at the same time to mimic high cell density

is to clone the cells onto a growth arrested ________________.

5) Cell separation method in which a cell type specific antibody raised against a cell surface

epitope is conjugated to the bottom of a petri dish is called ________________ .

6) After trypsinization if you add, medium _____ times the volume of trypsin used, it can

neutralize the toxic effect of trypsin.

7) FACS stands for ________________________________ .

8) Medium that has been used for the growth of other cells acquiring metabolites, growth factors
and matrix products from these cells are called ___________________________.

9) Once a primary culture is subcultured or passaged or transformed, it becomes known as a

_________

10) Cell lines derived from the _________ may contain more stem cells and precursor cells and
be capable of greater self-renewal than cultures from adults.
 KATHMANDU UNIVERSITY
End Semester Examination[C]
2014
Level : B. Tech. Course : BIOT 304
Year : III Semester : I
Time : 2 hrs 30 mins F. M. : 55
______________________________________________________________________________
SECTION”C”
(3Q 8 = 24)
Attempt any THREE questions.

1. List out the various techniques used in cell separation? Explain the working principle of
flow cytometry? [3+5]

2. Write a detailed flow chart for preparation of cell culture medium from powdered medium?
[8]

3. Describe the different physiochemical properties of medium used in cell culture? [8]

4. Discuss in detail the history of animal cell culture. [8]

SECTION ”D”

Attempt any SIX questions. (Question 7 is compulsory)

5. Explain the importance of bicarbonate buffer system in cell culture? [5]

6. Discuss the advantages of serum? [5]

7. What is dilution cloning, what should be considered while performing it in monolayer?

Explain how monolayer clones are isolated with diagram. [1+5]

8. What are the advantages of animal cell culture? [5]

9. How is primary culture cell isolated by cold trypsinization method? [5]

10. Write in brief about cell adhesion in animal cell culture. How does a suspension cell differ
from mono layer cells. [3+2]

SECTION ”E”

11. Write short notes on any TWO [2.52=5]

a) Haemocytometer

b) Cell preservation

c) Balanced salt solutions