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Cell Culture Techniques Questions and Answers

1.

The total number of live cells in a culture is counted using the frypan blue exclusion assay and is found to be 2.7 × 106 cells/ml. The culture is diluted 1 : 27 and then 100 μl seeded per well into a 96 well plate. What is the final cell density per well ?

Exam Year(s)

MH SET - 2018, SET Exam - 2018,

Answer: D

The final cell density is \(1 \times 10^{4}\) cells per well.

Given 2.7 × 106 live cells/ml, a 1 : 27 dilution, and 100 μl (0.1 ml) seeded per well.

\[ \begin{aligned} \frac{2.7 \times 10^{6}}{27} &= 1 \times 10^{5}\ \text{per ml} \\ 10^{5} \times 0.1 &= 1 \times 10^{4}\ \text{per well} \end{aligned} \]

\(1 \times 10^{5}\) is the concentration per millilitre, the answer if the 100 μl volume is forgotten. \(2.7 \times 10^{5}\) and \(2.7 \times 10^{4}\) skip the dilution. (The paper prints 'frypan blue'; the dye is trypan blue, which enters only dead cells.)

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2.

You are looking at a specimen with a compound microscope. The ocular lens has a magnification of 8X and the objective lens has a magnification of 2X. What is the total magnification ?

Exam Year(s)

MH SET - 2020, SET Exam - 2020,

Answer: D

The total magnification is 16X.

In a compound microscope the objective forms a magnified image, and the ocular (eyepiece) magnifies that image again, so the two magnifications multiply:

\[ 8 \times 2 = 16 \]

10X adds the two powers instead of multiplying them, and 8X and 2X are the powers of the single lenses on their own.

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3.

Cryopreservation is a process of :

Exam Year(s)

MH SET - 2021, SET Exam - 2021,

Answer: A

Cryopreservation is ex situ in vitro conservation.

In cryopreservation, seeds, embryos, pollen, shoot tips or cell cultures are stored at very low temperature, usually in liquid nitrogen at −196°C, where metabolism effectively stops. It takes material away from its natural habitat, so it is ex situ, and it keeps tissues and cells in culture vessels and vials rather than as whole growing plants, so it is in vitro.

Ex situ ex vitro conservation means botanical gardens, field gene banks and zoos. In situ conservation keeps organisms in their habitats, such as national parks and biosphere reserves, and is never in vitro.

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Answer: A

It retains many characteristics of the original tissue but has a limited lifespan. Once it is subcultured for the first time it becomes a cell line rather than a primary culture. The distinction matters because behaviour changes at that point.

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Answer: B

Cells are detached, diluted and reseeded before they exhaust the medium or fill the surface. Recording the passage number matters, since cells change with repeated subculture. A high-passage culture may behave quite differently from a low-passage one.

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Answer: C

The enzyme cleaves the proteins anchoring the cells to the surface and to one another. Exposure must be brief, since prolonged treatment damages surface proteins the cells need. Serum-containing medium is added afterwards to neutralise the enzyme.

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7.

State whether the following is true or false.
Trypsin should be left on adherent cells for as long as possible to ensure complete detachment.

Answer: D

Prolonged exposure digests surface proteins the cells depend on and reduces their viability. The usual practice is the shortest exposure that lifts the cells, followed by neutralisation with serum-containing medium. Longer is not better here, despite the intuition.

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Answer: A

Escaping the normal replicative limit lets the line be maintained indefinitely, which is convenient but means it differs from the tissue it came from. Such lines often carry chromosomal abnormalities. Convenience is bought at the cost of resemblance to the original tissue.

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Answer: B

Taken from Henrietta Lacks in 1951 without her knowledge or consent, it has been used in laboratories ever since. The circumstances of its origin remain an important case in research ethics. Her family received no acknowledgement or benefit for many decades.

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10.

State whether the following is true or false.
The HeLa cell line was established with the informed consent of the patient it came from.

Answer: C

The cells were taken without Henrietta Lacks's knowledge, and her family learned of their use only decades later. The case is now a standard teaching example in the ethics of human tissue. Consent practices in research changed considerably as a result.

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