Passage number in mammalian cells

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In summary, if a cell line has a maximum of 10 passages, freezing and thawing it would count as one passage. Therefore, if you have only done 4 passages and then froze the cells, there would be 5-6 passages left when thawed and passaged again. This method may not always work to regain lost characteristics in immortal cell lines, but some have tried re-passage through an animal to restore them.
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Goodie
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if a cell line has a maximum of 10 passages before it changes character and i only have done 4 passages of it and made the frozen stock of some of the cells. how many passages does this frozen stock have if i thaw it and passage it? will it be only 6 passages left or 10 passages?

thanks for any inputs.
 
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Typically you would consider the freeze-thaw another passage and call it around 5-6 passages left. I think the only way you can sometimes regain lost characteristics in immortal cell lines is to re-passage the cells back through an animal by creating a cell pellet and implanting it, but this may not work for all situations. Any one done this?
 
  • #3


Based on the information provided, the frozen stock would have 6 passages left if thawed and passed, assuming that the previous 4 passages were done correctly and there were no additional passages made from the frozen stock. It is important to keep track of the number of passages to ensure the integrity and consistency of the cell line. It is recommended to thaw and passage the cells within the remaining 6 passages to avoid any changes in character.
 

1. What is passage number in mammalian cells?

Passage number in mammalian cells refers to the number of times a cell culture has been subcultured or passaged. This is important for maintaining healthy and genetically stable cells in the laboratory.

2. Why is passage number important in cell culture?

The passage number of cells is important because it can affect the viability, growth rate, and genetic stability of the cells. As cells are passaged, they may accumulate mutations or changes in gene expression, which can alter their behavior and experimental results.

3. How often should I passage my mammalian cells?

The frequency of passaging cells depends on their growth rate and confluency. Generally, cells should be passaged when they reach 80-90% confluency to prevent overcrowding and nutrient depletion. This can range from every 2-3 days for fast-growing cells to every 7-10 days for slower-growing cells.

4. Can I use a high passage number of cells in my experiments?

It is generally recommended to use cells with a low passage number (less than 20) for experiments, as they are more likely to have stable genetic and phenotypic characteristics. However, some cell lines can be used at higher passage numbers with proper validation and controls.

5. How can I keep track of passage number in my cell culture?

To keep track of passage number, it is important to record each time the cells are split or subcultured. This can be done by labeling the cell culture vessel with the passage number, or by keeping a detailed record in a laboratory notebook. It is also helpful to freeze down cells at early passages and thaw a new vial periodically to prevent using cells with a high passage number.

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