EXPERIMENT: CRYOPRESERVATION OF CULTURED CELL LINES Based on the experiment, please answer the following questions. 1. Why cryoprotectant was used to freeze the cells? 2. Besides DMSO, suggest 3 other chemicals that potentially be used as cryoprotectant. 3. Describe the mechanism of DMSO in shielding the cells from extreme temperature. 4. In your opinion, why do we need to freeze the cells in high concentration? Justify your answer.

EXPERIMENT: CRYOPRESERVATION OF CULTURED CELL LINES Based on the experiment, please answer the following questions. 1. Why cryoprotectant was used to freeze the cells? 2. Besides DMSO, suggest 3 other chemicals that potentially be used as cryoprotectant. 3. Describe the mechanism of DMSO in shielding the cells from extreme temperature. 4. In your opinion, why do we need to freeze the cells in high concentration? Justify your answer.

Biomedical Instrumentation Systems

1st Edition

ISBN:9781133478294

Author:Chatterjee

Publisher:Chatterjee

Chapter24: New Technologies And Advances In Medical Instrumentation

Section: Chapter Questions

Problem 3P

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EXPERIMENT: CRYOPRESERVATION OF CULTURED CELL LINES Procedure for Cryopreserving MCF-7 and THP-1 Cells• Ensure that the culture is up to the late log phase (assumption is made based on the cell concentration). Prepare the cell suspension and determine the cell concentration.• Dilute the cell suspension in 1:1 ratio with freezing medium (final concentration approximately 1 X 106 – 1 X 107cells/ml is desired). • Dispense the cell suspension into prelabeled cryovial and place the vial in -80°C freezer overnight (temporary storage).• Transfer the cryovial into liquid nitrogen cryotank for long term storage the following day.• After day 3, determine the cell concentration in the freezing medium andcompare with that of the stock concentration. Question: Please briefly explain what is cryopreservation(1), how procedure above are useful to make this experiment successful(2)
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Experiment No. 1. Five rabbits were inoculated in the right lung and in the left side of the neck with five minims of sterilized water in which was suspended a sufficient quantity of a pure culture (third generation) of the tubercle bacillus to render the liquid quite perceptibly turbid The needle of the Koch’s inoculating syringe was inserted subcutaneously on the left side of the neck and in the third intercostal space to a depth of thirty millimetres on the right side. These animals were then confined in a small box and put in a dark cellar. They were thus deprived of light, fresh air and exercise and were also stinted in the quantity of food given them while being themselves artificially infected with the tubercle bacillus. Experiment No. 2. Five healthy rabbits were placed under the following conditions: A fresh hole about ten feet deep was dug in the middle of a field, and the animals having been confined in a small box with high sides but no top, were lowered to the bottom of…
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