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Storage in incubator?

  1. Mar 14, 2005 #1
    i wonder how long can i store my cells in incubator if i don't need the cells right away, but in coming days? should i change the medium regularly when i store the cells in the incubator instead of freezing? or do i have to freeze down the cells immediately and thaw them when i need them?

    appreciate for all help.
     
  2. jcsd
  3. Mar 15, 2005 #2

    DocToxyn

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    If it's days you are considering, it's probably best to keep the culture going in the incubator. Split the cultures at a dilution that will allow them to achieve the density you desire on or near the day you plan to use them.

    As far as feeding, it usually depends on the cell type, which dictates how metabolically active they are. As an example of cells I've maintained, PC12 cells (from a pheochromocytoma) require feeding every other day, but Hepa 1C1C7 cells (mouse liver) can go about four days between feedings. The state of the culture can also determine how often you feed, thus a fully confluent monolayer might require more frequent feedings than a very lightly populated flask that was just seeded. If you can't find the information on your specific cell type in the literature, your best bet would be to include phenol red in your medium. This indicator will turn from ~cherry red to yellow as the cells metabolize and cast off acidic waste products. If the medium starts to look yellow, it's best to do a medium change.

    Having said this, if you don't already have a good supply of frozen stocks to work from, it would be best for you to freeze some away for future use, you never know when your cultures might get contaminated and require starting over. Another thing to consider is the passage number, or how many times you have split the cells. Sometimes certain characteristics in a cell line can change as they "age" so keeping track of the passages can aid in limiting/controlling this potential variation. Many researchers will describe passage number used for their experiments so that others are aware of the state of the cells.
    appreciate for all help.
     
  4. Mar 15, 2005 #3
    Thanks alot. :smile:
     
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