General procedure for cell culture MCQs With Answer

Introduction:
General procedure for cell culture MCQs With Answer is an essential study aid for B. Pharm students preparing for practical exams and competitive tests. This concise, keyword-rich guide covers core topics such as aseptic technique, sterile media preparation, passaging, cryopreservation, contamination control, cell counting, incubator settings and primary culture versus established cell lines. Focused on good cell culture practice (GCCP), it explains sterilization methods, biosafety cabinet use, serum and serum-free media considerations, and troubleshooting common problems. Clear, exam-oriented MCQs reinforce concepts and improve retention. Now let’s test your knowledge with 50 MCQs on this topic.

Q1. What is the primary purpose of using a biosafety cabinet (Class II) during cell culture?

To maintain sterile working area and protect operator and samples
To incubate cell cultures at 37°C
To centrifuge cell suspensions safely
To store cell culture reagents

Correct Answer: To maintain sterile working area and protect operator and samples

Q2. Which reagent is commonly used to detach adherent mammalian cells from the culture flask?

Trypsin-EDTA solution
Phosphate buffered saline (PBS)
Fetal bovine serum (FBS)
Dimethyl sulfoxide (DMSO)

Correct Answer: Trypsin-EDTA solution

Q3. What is the role of fetal bovine serum (FBS) in cell culture media?

Provides growth factors, hormones and attachment factors
Acts as an antibiotic to prevent contamination
Neutralizes trypsin after cell detachment
Serves as a cryoprotectant during freezing

Correct Answer: Provides growth factors, hormones and attachment factors

Q4. Which method is best for long-term storage of mammalian cell lines?

Cryopreservation in liquid nitrogen with DMSO
Room temperature storage in flasks
Storage at 4°C in the incubator
Frequent passaging at high confluency

Correct Answer: Cryopreservation in liquid nitrogen with DMSO

Q5. What does confluency refer to in adherent cell culture?

Percentage of surface area covered by cells
Number of cells per milliliter in suspension
pH of the culture medium
Amount of serum in the medium

Correct Answer: Percentage of surface area covered by cells

Q6. Which indicator in culture medium helps monitor pH changes during cell culture?

Phenol red
Trypan blue
EDTA
Glucose

Correct Answer: Phenol red

Q7. What is the main purpose of using antibiotics in cell culture media?

To reduce risk of bacterial contamination during routine culture
To enhance cell proliferation rate
To promote cell differentiation
To act as a cryoprotectant

Correct Answer: To reduce risk of bacterial contamination during routine culture

Q8. Which of the following is a common sign of mycoplasma contamination?

Slow growth and altered cell morphology without visible particulates
Large floating clumps of bacteria visible under light microscope
Immediate turbidity in culture medium within hours
Intense foaming of the culture medium

Correct Answer: Slow growth and altered cell morphology without visible particulates

Q9. Which temperature is most commonly used for culturing human-derived mammalian cells?

37°C
25°C
4°C
50°C

Correct Answer: 37°C

Q10. What is the purpose of trypan blue exclusion test?

To assess cell viability by membrane integrity
To sterilize cell suspensions
To enhance cell attachment to culture flasks
To fix cells for microscopy

Correct Answer: To assess cell viability by membrane integrity

Q11. What is the correct action if you notice sudden turbidity and foul odor in a culture flask?

Discard the culture as contaminated following biosafety procedures
Add antibiotics to salvage the culture
Increase incubation temperature to kill contaminants
Transfer cells to a new flask immediately

Correct Answer: Discard the culture as contaminated following biosafety procedures

Q12. Which centrifuge parameter is most appropriate to pellet mammalian cells gently?

Low speed (e.g., 200–500 x g) for short time
High speed (e.g., 10,000 x g) for a long time
Maximum speed of the rotor for a very short time
0 x g without spinning

Correct Answer: Low speed (e.g., 200–500 x g) for short time

Q13. What does “passaging” or “subculturing” mean?

Transferring cells to fresh medium to prevent over-confluency
Freezing cells for long-term storage
Counting cells using a hemocytometer
Measuring cell viability with dyes

Correct Answer: Transferring cells to fresh medium to prevent over-confluency

Q14. Which vessel is commonly used for routine adherent cell culture in labs?

T-flask
Erlenmeyer flask without surface treatment
Glass beaker
Petri dish used for bacterial culture only

Correct Answer: T-flask

Q15. Which of the following best describes a primary culture?

Cells directly isolated from tissue and maintained for limited passages
Immortalized cell line that grows indefinitely
Cell-free extract used for biochemical assays
Cells genetically modified to express a marker

Correct Answer: Cells directly isolated from tissue and maintained for limited passages

Q16. What is the function of EDTA in trypsin-EDTA solution?

Chelates calcium and magnesium to help cell detachment
Acts as a pH indicator
Serves as an antibiotic agent
Preserves cells during freezing

Correct Answer: Chelates calcium and magnesium to help cell detachment

Q17. Which practice reduces cross-contamination between cell lines?

Using dedicated media and pipettes for each cell line
Sharing the same hood for multiple lines at once
Using antibiotics to mask contamination signs
Labeling flasks with only date, not cell name

Correct Answer: Using dedicated media and pipettes for each cell line

Q18. Which assay is used to determine cell proliferation rate and doubling time?

Growth curve plotting cell counts over time
Gram staining of culture medium
Mycoplasma PCR only
Trypan blue staining for viability at a single time point

Correct Answer: Growth curve plotting cell counts over time

Q19. What is a common cryoprotectant used to prevent ice crystal formation during freezing?

Dimethyl sulfoxide (DMSO)
Fetal bovine serum (FBS)
EDTA
Phenol red

Correct Answer: Dimethyl sulfoxide (DMSO)

Q20. Which parameter is critical to control in a CO2 incubator for mammalian cell culture?

CO2 concentration, temperature and humidity
Light intensity inside the incubator
Rotation speed of the incubator
Oxygen concentration at 100% only

Correct Answer: CO2 concentration, temperature and humidity

Q21. What does sterility testing of culture medium typically check for?

Bacterial and fungal contamination in prepared media
Cell viability after thawing
Confluency of adherent cultures
Genetic stability of cell lines

Correct Answer: Bacterial and fungal contamination in prepared media

Q22. Which is a sign that adherent cells are ready to be passaged?

Cells reach 70–90% confluency with healthy morphology
Medium turns pink due to phenol red
Cells detach completely without treatment
Culture shows visible bacterial colonies

Correct Answer: Cells reach 70–90% confluency with healthy morphology

Q23. What is the main disadvantage of routine use of antibiotics in cell culture?

It can mask low-level contamination and promote resistant strains
It increases cell differentiation speed
It acts as a strong growth stimulant
It prevents cryopreservation success

Correct Answer: It can mask low-level contamination and promote resistant strains

Q24. Which test is commonly used to detect mycoplasma contamination?

PCR-based assays or specific mycoplasma detection kits
Gram staining of culture medium
Trypan blue exclusion test
Measuring phenol red color only

Correct Answer: PCR-based assays or specific mycoplasma detection kits

Q25. What is the ideal action when thawing frozen cells for culture?

Thaw quickly in a 37°C water bath and dilute promptly in warm medium
Thaw slowly at room temperature overnight
Leave cryovials at -20°C to equilibrate then thaw
Thaw directly on ice to slow warming

Correct Answer: Thaw quickly in a 37°C water bath and dilute promptly in warm medium

Q26. Which factor primarily determines whether cells grow as adherent or suspension cultures?

Cell type and anchorage dependence
Concentration of antibiotics
Color of the culture medium
Amount of DMSO in the medium

Correct Answer: Cell type and anchorage dependence

Q27. What is the main purpose of performing cell authentication (e.g., STR profiling)?

To confirm the identity of a cell line and prevent misidentification
To sterilize the cell culture before experiments
To count cells accurately
To increase cell proliferation rate

Correct Answer: To confirm the identity of a cell line and prevent misidentification

Q28. Which media component supplies energy and carbon source for cultured cells?

Glucose
Phenol red
Trypsin
Dimethyl sulfoxide

Correct Answer: Glucose

Q29. What is the recommended split ratio for routine passaging to maintain exponential growth?

Depends on cell line; common ratios range from 1:2 to 1:10
Always 1:100 regardless of cell type
0.5:1 to increase confluency quickly
Never split; always keep original flask

Correct Answer: Depends on cell line; common ratios range from 1:2 to 1:10

Q30. Which of the following is NOT a sterile technique in cell culture?

Touching media opening with gloved hand inside the hood
Flaming the neck of a bottle before opening (where appropriate)
Working quickly and minimizing exposure of sterile surfaces
Using sterile consumables and filtered pipette tips

Correct Answer: Touching media opening with gloved hand inside the hood

Q31. Why is it important to equilibrate culture media in the incubator before use?

To stabilize temperature and CO2-dependent pH
To sterilize the medium further
To increase serum concentration
To remove dissolved oxygen completely

Correct Answer: To stabilize temperature and CO2-dependent pH

Q32. Which practice improves reproducibility in cell culture experiments?

Using consistent cell passage number and standardized protocols
Changing media composition randomly between experiments
Varying incubation temperatures deliberately each run
Using different suppliers for the same reagent each day

Correct Answer: Using consistent cell passage number and standardized protocols

Q33. What is anchorage dependence?

Requirement of certain cells to attach to a surface to grow
Ability of cells to grow in suspension without attachment
Requirement of CO2 for cell survival
Dependence on antibiotics to thrive

Correct Answer: Requirement of certain cells to attach to a surface to grow

Q34. Which of the following is a common method to decontaminate small spills of live cell media inside the hood?

Wipe with 70% ethanol followed by disinfectant and absorbent towels
Leave the spill to dry under UV light only
Flush with sterile water and resume work
Cover with paper and continue working

Correct Answer: Wipe with 70% ethanol followed by disinfectant and absorbent towels

Q35. What is the purpose of serum-free media formulations?

To reduce variability and control defined growth conditions
To increase bacterial contamination risk
To ensure cells cannot adhere to surfaces
To act primarily as cryoprotectants

Correct Answer: To reduce variability and control defined growth conditions

Q36. Which measurement requires use of a hemocytometer?

Total cell count per unit volume
pH value of medium
CO2 concentration in incubator
Serum concentration in medium

Correct Answer: Total cell count per unit volume

Q37. Which of the following best describes cross-contamination of cell lines?

Unintended mixing of different cell lines leading to misidentification
Intended co-culture of two cell types for an experiment
Contamination by bacteria only
Freezing cells in multiple vials

Correct Answer: Unintended mixing of different cell lines leading to misidentification

Q38. What should be done before opening sterile consumables in a biosafety cabinet?

Wipe surfaces with 70% ethanol and allow airflow to stabilize
Turn off cabinet airflow to prevent turbulence
Expose consumables to UV light for 24 hours without wiping
Open items outside the hood to save time

Correct Answer: Wipe surfaces with 70% ethanol and allow airflow to stabilize

Q39. Which step neutralizes trypsin after cell detachment?

Adding serum-containing medium
Washing with sterile water
Adding DMSO
Increasing incubator CO2

Correct Answer: Adding serum-containing medium

Q40. Why is documentation and labeling important in a cell culture lab?

Ensures traceability, prevents misidentification and supports reproducibility
Reduces the need for sterile technique
Allows mixing of different cell lines without record
Replaces the need for regular mycoplasma testing

Correct Answer: Ensures traceability, prevents misidentification and supports reproducibility

Q41. What is the effect of prolonged over-confluency on cell cultures?

Altered cell physiology, contact inhibition, and potential differentiation
Improved cell viability and faster proliferation
Immediate sterilization of the culture
Reduced need for passaging indefinitely

Correct Answer: Altered cell physiology, contact inhibition, and potential differentiation

Q42. Which vessel surface treatment promotes attachment of adherent mammalian cells?

Tissue culture-treated surface
Non-treated glass surface excluding treatment
Hydrophobic plastic without treatment
Uncoated silicone

Correct Answer: Tissue culture-treated surface

Q43. What does “GCCP” stand for in cell culture practice?

Good Cell Culture Practice
General Clinical Cell Procedure
Global Cell Contamination Protocol
Guided Cell Counting Practice

Correct Answer: Good Cell Culture Practice

Q44. Which is a common immediate action after accidental exposure of skin to culture reagents?

Wash the area thoroughly with soap and water and report the incident
Wipe with a tissue and ignore
Cover the area with a bandage without washing
Apply disinfectant intended for surfaces directly on skin

Correct Answer: Wash the area thoroughly with soap and water and report the incident

Q45. Which type of contamination often shows visible filamentous growth or fuzzy colonies in culture?

Fungal contamination
Mycoplasma contamination
Viral contamination
Cell line cross-contamination

Correct Answer: Fungal contamination

Q46. Which practice helps minimize foam formation and shear stress in suspension cultures?

Use gentle agitation and appropriate baffled vessels or anti-foaming agents
Increase agitation speed to maximum
Add high concentrations of serum to reduce shear
Shake cultures vigorously by hand

Correct Answer: Use gentle agitation and appropriate baffled vessels or anti-foaming agents

Q47. Why perform routine mycoplasma testing on cell cultures?

Because mycoplasma alters cell behavior and experimental results
Because mycoplasma improves cell growth
Because mycoplasma only affects bacteria
Because mycoplasma makes cultures visibly turbid immediately

Correct Answer: Because mycoplasma alters cell behavior and experimental results

Q48. What is the recommended composition of a basic complete growth medium?

Basal medium with salts, glucose, amino acids, vitamins and serum or defined supplements
Pure distilled water only
DMSO with antibiotics only
Phenol red diluted in ethanol

Correct Answer: Basal medium with salts, glucose, amino acids, vitamins and serum or defined supplements

Q49. What is the first step before entering a cell culture room to work?

Put on appropriate PPE (lab coat, gloves, and eye protection) and disinfect hands
Open all incubator doors to ventilate
Start handling cultures immediately without preparation
Bring food and drink into the room

Correct Answer: Put on appropriate PPE (lab coat, gloves, and eye protection) and disinfect hands

Q50. Which practice helps preserve the genetic stability of cultured cell lines?

Limiting passage number and using early-passage stocks from authenticated seed vials
Continuously passaging cells indefinitely without freezing stocks
Using random media changes with unknown supplements
Increasing DMSO concentration during regular culture

Correct Answer: Limiting passage number and using early-passage stocks from authenticated seed vials

G S Sachin

I am a Registered Pharmacist under the Pharmacy Act, 1948, and the founder of PharmacyFreak.com. I hold a Bachelor of Pharmacy degree from Rungta College of Pharmaceutical Science and Research. With a strong academic foundation and practical knowledge, I am committed to providing accurate, easy-to-understand content to support pharmacy students and professionals. My aim is to make complex pharmaceutical concepts accessible and useful for real-world application.

Mail- Sachin@pharmacyfreak.com

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