Culturing and Harvesting Techniques Flashcards
(91 cards)
1
Q
The following enzymes can be used to dissociate Chorionic villus Samples (CVS) EXCEPT FOR: A. RNAse B. Collagenase C. Trypsin D. Protease
A
A
2
Q
All of the following have buffering capability EXCEPT:
A. Sodium bicarbonate B. L-Glutamine C. Hepes D. FCS
A
B
3
Q
What type of media is necessary for the culture of Fragile X cases?
A. Media with extra FCS B. Folic acid-deficient media C. Complete media D. Lysine-rich media
A
B
4
Q
The most important reason why Biological Safety Cabinets are used instead of Horizontal
Laminar Flow Hoods in the handling of human tissues is
A. To provide a clean working area. B. To prevent contamination of the specimen. C. To protect the technologist who is processing the specimen. D. To prevent cross-contamination between two different cell lines.
A
C
5
Q
In which of the following situations would one suspect bacterial contamination?
A. The culture media is red-orange. B. The culture had a dark mass floating on the surface of the media. C. The culture media is turbid in appearance. D. The pH of the media is 7.2.
A
C
6
Q
Which of the following can be used for serial culture from amniotic fluid?
A. Erythrocyte B. Lymphocyte C. Epithelial cells D. Fibroblast
A
D
7
Q
Which of the following chemicals is used for cryopreservation?
A. Methanol B. Ethyl alcohol C. Dimethyl sulfoxide (DMSO) D. Sodium hyposchorite
A
C
8
Q
Adding the drug Colcemid to cell cultures can cause all of the following phenomenon to
occur EXCEPT:
A. Chromosome condensation B. Increase in the number of metaphases C. Cell swelling D. Depolymerization of the spindle apparatus
A
C
9
Q
During slidemaking all the following factors are important EXCEPT FOR:
A. Drying rate (evaporation)
B. Quality of the glass slides C. Slides are first dipped in water D. Slides are first dipped in acetic acid
A
D
10
Q
A good banding pattern will be expected when we observe chromosomes before banding
under the phase scope with the following appearance:
A. Dark
B. Pale C. Shiny D. Separated chromatids
A
A
11
Q
Which of the following factors is not important for chromosome spreading?
A. Hypotonic treatment time B. Dropping the cell from an increased height C. Cell suspension concentration D. Mitotic index
A
D
12
Q
If short chromosomes result after harvesting, what change may alleviate this condition for the next harvest?
A. increase concentration of methanol in fixative B. decrease exposure to hypotonic solution C. decrease concentration of Colcemid D. increase concentration of Colcemid
A
C
13
Q
When slide making, what appearance would the technologist prefer the chromosome to have if the slides will be G-banded?
A. Gray
B. Refractile C. very black D. chromatids separated
A
A
14
Q
Which of the following apply to mitotic arrestants?
A. Result in depolymerization of the mitotic spindle B. Colchicine C. Colcemid D. all of the above
A
D
15
Q
Which of the following apply to hypotonic solutions?
A. Are responsible for lysing red blood cells B. Cause cells to swell C. 0.075M KCL D. all of the above
A
D
16
Q
When dropping slides, the majority of chromosomes appear to be short. This can result from which of the following?
A. A long exposure to Colcemid
B. A short exposure to Colcemid C. A short exposure to hypotonic D. Improper fixation
A
A
17
Q
After what period of time do PHA peripheral blood lymphocytes reach their first division?
A. 12 hours B. 24 hours C. 36 hours D. 48 hours
A
D
18
Q
Which of the following is information that is important to consider when freezing cells?
A. age of the culture B. concentration of cells C. cell type D. media used
A
B
19
Q
24 BM and 72 BM LPS cultures would be set up on a new CLL with a wbc of 5 x 106.
A. True .
B. False .
A
TRUE
20
Q
Which of the following may cause cell death?
A. A five percent C02 atmosphere B. A 24 hour delay in culturing C. A pH of 7.2 D. Incubation at a temperature of 40oC
A
D
21
Q
Which of the following pH will provide optimal cell growth?
A. 6.8-7.0 B. 7.0-7.2 C. 7.2-7.4 D. 7.4-7.6
A
C
22
Q
Which of the following tissues grows most slowly in culture?
A. fetal tissue B. skin C. gonad D. amniotic fluid cells
A
B
23
Q
Which of the following media supplements is optional for long-term cultures?
A. insulin B. antibiotics antifungicides C. growth factors D. fetal calf serum
A
B
24
Q
Which of the following best describes a media that has a low serum level?
A. defined B. maintenance C. growth D. complete
A
B
25
The statements below refer to culturing methods. Which statement is FALSE?
A. The preferred mode for cytogenetic analysis of solid tumors is long-term culture.
B. The success rate of cytogenetic analysis of solid tumors is less than that of leukemias currently.
C. Culture of epithelial tumors is more difficult than other types of tumors.
D. A Biosafety Cabinet is required for tissue culture.
A
26
Regarding the following statements, which is FALSE?
A. Epithelial cells cannot be morphologically distinguished from mesenchymal cells in culture when viewed
under an inverted phase contrast microscope.
B. Contamination with normal fibroblasts can give an erroneous result.
C. Different media are used in the culture of different tumor types.
D. all of the above
A
27
Harvest of a 14-day-old amniotic fluid culture results in many cells, but few mitoses. The most likely cause of the
low mitotic rate is:
A. Senility of the culture.
B. Chromosome abnormality
C. Maternal age
D. Contact inhibition due to high cell confluence
D
28
The culture method used to obtain chromosome preparations from amniotic fluid is considered:
A. Short term culture
B. Intermediate term culture
C. Long term
D. None of the above
A
29
The fixative step of cell harvesting does not:
A. Kill the cell by fixing the cell membrane
B. Remove water from the cells
C. Extract nuclear histones
D. Disperse the chromosomes
D
30
Which of the following is NOT a mitotic inhibitor?
A. Colcemid
B. Vinblastine
C. Ethidium bromide
D. Colchicine
C
31
A hypotonic solution is:
A. A solution with a higher salt concentration than that of cytoplasm.
B. A solution with a lower salt concentration than that of cytoplasm.
C. A solution with an equal salt concentration as that of cytoplasm.
D. A solution used to wash cells.
B
32
Carnoy's Fixative is the standard fixative used in chromosome harvesting procedures. Which of the following is a
description of this solution?
A. 3 parts glacial acetic acid to 1 part methanol
B. 1 part glacial acetic acid to 2 parts methanol
C. 2 parts glacial acetic acid to 1 part methanol
D. 1 part glacial acetic acid to 3 parts methanol
D
33
If the metaphase portion of the cell cycle is 15 minutes and a cytogeneticist chooses to treat a peripheral blood specimen with colcemid for 45 minutes, what is the increase in mitotic cells, which will be observed after the harvest procedure is completed?
A. 100%
B. 200%
C. 300%
D. 400%
C
34
When newly dropped slides are observed, the chromosomes appear to be nonuniform. The chromosomes vary in quality not only within the cell but also across the slide. Which of the following would cause this unevenness?
A. Dirty slides
B. Non-uniform drying of the slide
C. Air currents
D. All of the above
D
35
What is the concentration of KCL (hypotonic solution) most often used for harvesting?
A. 0.04 M
B. 0.06 M
C. 0.075 M
D. 0.75 M
C
36
How many grams of potassium chloride is added to 1000 ml of double deionized water to make 0.075M KCL?
A. 5.58 g
B. 2.79 g
C. 4.79 g
D. 7.5 g
A
37
What do mitotic arrestants prevent cells from entering?
A. S phase
B. Prophase
C. Anaphase
D. Telophase
C
38
A technologist is having a problem with cytoplasm remaining around the cells. Two extra fixative steps have been added, but the problem persists and the cell pellet is white and fluffy in appearance. What else can this technologist do to alleviate this problem?
A. Manipulate the drying time
B. Decrease the methanol concentration
C. Wash two more times with fixative
D. Drop the cell suspension onto the slide from a lesser height
A
39
Which of the following factors concerning hypotonic solutions DO NOT affect chromosome spreading?
A. Concentration of the hypotonic solution
B. Temperature of the hypotonic solution.
C. Time period cells are exposed to the hypotonic solution.
D. Number of washes with hypotonic.
D
40
Disadvantages of the direct harvest method as opposed to harvesting cultured cells for CVS analysis are:
A. Poor quality chromosomes and increased pseudomosaicism.
B. Increased incidence of maternal cell contamination
C. Requires fluorescent marker studies
D. Increased pseudomosaicisim
A
41
Cytogenetic studies have been requested for a patient with a diagnosis of questionable leukemia. This patient has a low white count, circulating blasts, and has received chemotherapy and a blood transfusion. Which of the following procedures is LEAST likely to be informative?
A. Direct harvest of bone marrow
B. Unstimulated culture of peripheral blood lymphocytes
C. PHA stimulated blood culture
D. Unstimulated culture of washed blood cells
C
42
PHA is a common additive to blood cultures. What category of additive is it?
A. a mitogen
B. an anticoagulant
C. growth serum
D. Arresting solution
A
43
On Monday morning the technologist noticed widespread cell death in incubator A. He checked the environment and conditions of the cultures. What condition would likely be the cause of the cell death?
A. 5% C02 atmosphere
B. 40oC incubator temperature
C. a pH of 7.3
D. all conditions would cause cell death
B
44
Which media component is unstable?
A. Serum
B. L-glutamine
C. Antibiotics
D. amino acids
B
45
What is the purpose of establishing a PHA-stimulated blood culture for a leukemia patient?
A. To check for circulating immature cells in the peripheral blood.
B. To see if an abnormality is constitutional rather than acquired
C. To look for early rearrangements which may have predisposed this individual to develop cancer.
D. To study leukemic cells of T-cell origin
B
46
What is the purpose of establishing an unstimulated blood culture for a leukemia patient?
A. To check for circulating immature cells in the peripheral blood.
B. To see if an abnormality is constitutional rather than acquired.
C. To look for early rearrangements which may have predisposed this individual to develop cancer.
D. all of the above
A
47
PHA will affect which of the following types of cells?
A. B-cells
B. Platelets
C. T-cells
D. Phagocytes
C
48
Which of the following is a characteristic of the vertical but not horizontal laminar flow hood?
A. Protects the culture from contamination
B. Should not be used with transformed cell lines.
C. Protects the worker from contamination
D. Sterlizes the air entering the hood
C
49
The statements below refer to culturing methods. Which statement is FALSE:
A. The preferred mode for cytogenetic analysis of solid tumors is long term culture.
B. The success rate of cytogenetic analysis of solid tumors is less than that of leukemias currently.
C. Culture of epithelial tumors is more difficult than other types of tumors.
D. A Biosafety Cabinet is required for tissue culture.
A
50
This type of culture has a decreased turn around time and allows easier distinction between mosaicism and pseudomosaicism.
A. Primary amniotic fluid cell culture
B. In situ-amniotic fluid culture
C. Flask-amniotic fluid culture
D. Short term amniotic fluid culture
B
51
If a laboratory is consistently having problems with poor culture growth, what measures could remedy this situation?
A. Test for mycoplasma contamination
B. Increase temperature to 38.5oC
C. Add trypsin
D. Increase antibiotic concentration
A
52
One technologist experienced culture failure on all the cultures she set-up on Wednesday afternoon; all the
cultures set-up by her counterpart were progressing well. What is the possible cause of this catastrophe?
A. Microbial contamination
B. Toxic culture vessels
C. 20% growth serum in media
D. Cells cultured 30 hours after collection.
A
53
The lab supervisor detected contamination in a 5 day old in situ amniotic fluid culture. This was one of four
cultures set-up on this patient. What would be the best course of action in this situation?
A. Increase antibiotic concentration
B. Refeed the culture
C. Transfer the specimen to another culture vessel
D. Harvest immediately
D
54
How would a technologist know when to harvest an in situ amniotic fluid culture?
A. By the number of days in culture
B. Within 12 hours of subculture
C. The number of colonies present
D. Two of these
C
55
The major difference between the in situ and flask methods lies in the harvest technique.
A. True .
B. False .
A
56
Monday morning around 2:30, Incubator A ran out of carbon dioxide. What change in the cultures, if any would
you expect for the technologists to find at 8:00 am Monday.
A. About the same pH
B. A lower pH
C. A higher pH
D. A moderate pH
C
57
Culture reagents and chemicals may be used without adverse effects after they have expired.
A. True .
B. False .
B
58
Amniotic fluid cultures are generally started in a medium with what percentage of serum.
A. 5%
B. 10%
C. 15%
D. 20%
B
59
PHA
A. Stimulates the large lymphocytes
B. Transforms polymorphonuclear leukocytes
C. Causes decrease in RNA activity
D. Causes peak in DNA synthesis at 72 hours
E. Is needed throughout cell cycle to produce sufficient metaphases
D
60
PHA
```
A. Swells cells
B. Removes water from cells
C. Stimulates mitotic activity
D. Precipitates nucleic acid
E. Prevents clotting
```
C
61
In short term culture the following is/are true:
A. Incubation in a CO2 atmosphere is essential.
B. Most commercially prepared media is suitable
C. Plasticware is never toxic to cells.
D. All of the above.
B
62
"Starring-in" is a method of
A. Obtaining the specimen from the patient
B. Mincing the tissue
C. Immobilizing the tissue in the culture vessel
D. Fixing the cells
E. Preparing the slides
C
63
A change of 1 pH unit in culture medium is caused by a ________ change in CO2 concentration.
```
A. 10-fold
B. 25-fold
C. 50-fold
D. 100-fold
E. 1000-fold
```
A
64
Five percent CO2 results in a pH of
```
A. 6.5
B. 6.8
C. 7.0
D. 7.4
E. 7.8
```
D
65
What is the most commonly used growth supplement for culture medium?
A. Insulin
B. Tranferrin
C. L-glutamine
D. Fetal bovine serum
D
66
Which of the following biological safety cabinets are recommended for use in cytogenetic laboratories?
A. Biological Safety Cabinet Class I
B. Biological Safety Cabinet Class II
C. Biological Safety Cabinet Class III
D. All of the above
B
67
When subculturing a long-term cell line, a technician finds the cells are not easily removed by their trypsin
solution. What can be added to the trypsin to increase its effectiveness in removing the cells from the flask?
A. Serum
B. Calcium and magnesium
C. EDTA
D. L-glutamine
C
68
Which of the following microorganisms can adversely effect long-term cultures?
A. Mycoplasma
B. Bacteria
C. Fungus
D. All of the above
D
69
Which of the following is NOT an advantage of tissue culture (in vitro) studies over patient studies (in vivo)?
A. Less expensive
B. More ethical
C. Possibility of contamination
D. Can directly target cells with test agent
C
70
What microscope is used to view living monolayer cell cultures?
A. Brightfield microscope
B. Fluorescent microscope
C. Stereomicroscope
D. Inverted phase microscope
D
71
Unstimulated blood cultures grow out of what type of cells?
A. Segmented neutrophils
B. Lymphocytes
C. Blast forms
D. Monocytes
C
72
Stimulated blood cultures grow out what type of cells?
A. Basophils
B. Lymphocytes
C. Immature forms
B
73
What is NOT a principle behind the culturing of solid tumors?
A. Culturing of living cells provides better morphology chromosomes for analysis than chromosomes obtained by direct harvest.
B. To generate an expandable cell population which may be used for sequential and long-term experimentation.
C. To determine which chromosomal rearrangements are due to in vitro artifacts vs. those which represent the true in vivo condition.
D. None of the above are principles behind the culturing of solid tumors.
C
74
Collagenase is used in the analysis of solid tumors to:
A. Dissociate cells
B. To supplement culture media
C. To prevent mycoplasma contamination
D. To subculture the monolayer cultures.
A
75
The inverted microscope is essential in the monitoring of cell cultures by allowing the technologist to visualize all
but which of the following cellular features?
A. Cellular confluency
B. Chromosomal abnormalities
C. Mitotic cells
D. Cellular attachment
B
76
The next day after a solid tumor specimen has been set up in culture the tissue culturist notices that the culture medium has turned a bright yellow color. When the flask is observed under the inverted microscope, a large number of small dot shaped structures are observed. What is the most likely cause of this pH change and the appearance of these structures?
A. The specimen was received from the necrotic center of the tumor.
B. The specimen was not collected under sterile conditions.
C. The specimen was not grown in the appropriate media.
D. The specimen was not transported at the appropriate temperature.
B
77
Which of the following is not an effect of fixation?
A. Remove water from the cells
B. A short exposure to Colcemid
C. A short exposure to hypotonic
D. Improper fixation
B
78
Upon what cell structure does Colchicine have its effect?
A. Centrioles
B. Spindle fibers
C. Centromeres
D. Chromosomes
B
79
This chemical is often used to help swell cells in the harvest procedure.
A. Thymidine
B. KCL
C. Colchicine
D. Ethidium bromide
B
80
This chemical is more toxic than its more recent synthetic counterpart.
A. Thymidine
B. KCL
C. Colchicine
D. Ethidium bromide
C
81
This chemical is used to synchronize cells.
A. Thymidine
B. KCL
C. Colchicine
D. Ethidium bromide
A
82
This chemical is used as a chromosome elongating agent.
A. Amethopterin
B. KCL
C. Colchicine
D. Ethidium bromide
D
83
Which of the following is not a mitotic stimulant?
A. PHA
B. Citrate
C. Pokeweed
D. LPS
B
84
Carnoy's fixative:
A. Swells cells
B. Stops hypotonic action
C. Stimulates mitotic activity
D. Prevents clogging
B
85
Which of the following chemicals is NOT an intercalating agent?
A. Actinomycin D
B. Ethidium bromide
C. Methotrexate (Amethopterin)
D. Quinacrine dihydrochloride
C
86
This cell cycle blocking agent causes cells to be arrested in the S-phase of the cell cycle through the inhibition of
dihydrofolate reductase.
A. Methotrexate (Amethopterin)
B. 5-Bromodeoxyuridine
C. Ethidium bromide
D. Bleomycin
A
87
This chemical is a thymidine analog that replaces thymidine easily in mammalian cells.
A. 5-Fluorodeoxyuridine
B. Adenine
C. 5-Bromodeoxyuridine
D. Hoechst 33258
C
88
Which of the following environmental factors can have an impact on drying time and therefore impact
chromosome spreading during slide making?
A. Temperature
B. Humidity
C. Air currents
D. All of the above
D
89
A new hypotonic solution causes overspreading of chromosomes. What technique can be used to prevent the
overspreading?
A. Drop cells on a wet slide.
B. Increase the height from which cells are dropped.
C. Decrease the cell concentration
D. Reduce the drying time
D
90
The hypotonic treatment of cells during the harvesting procedure is used to:
A. Remove water from the cells
B. Disperse the chromosomes
C. Enhance chromosome morphology
D. Remove the H1 fraction of histones
B
91
All of the following may be used for a hypotonic during the harvest procedure EXCEPT:
A. Methanol
B. Sodium citrate
C. Water
D. Potassium chloride
A
