Addition of cryoprotectant in freezing cell (like DMSO) is missing.
@celsojoven3525
3 жыл бұрын
Umihealth soutwoods
@breadfan262
Жыл бұрын
Freezing medium contains DMSO
@insanity.defeated
Жыл бұрын
Isn't it usually 90% fbs 10% dmso?
@pashantanu
Жыл бұрын
Depends on the cell line, the freezing procedure are specifically mentioned in spec-sheet provided by ATCC after purchase. Normally in secondary cultures, we freeze in freezing media containing 5% DMSO in working media (10%FBS+plain media+supplements (if any)).
@katarinamajerikbehinska3850
10 ай бұрын
Thank you for this video series. It helps so much :)
@thermofisher
10 ай бұрын
Hello katarinamajerikbehinska, Glad it was helpful!
@lissethsilva7461
3 жыл бұрын
How do you know when your cells are ready to be frozen? Is it at a specific passage number?
@metallicamadsam
Жыл бұрын
the earlier the passage the better. if you thaw a cell type with a low passage number, and that cell stock is less than 2, then you make more aliquots of that low passage number cell stock
@kosheeka
3 ай бұрын
Passage number alone isn't the only indicator for freezing cells in culture. Here's what matters most: • Cell health and viability: Cells should be actively growing, with high viability, usually >90%. (You can look for minimal cell death and signs of stress.) • Exponential growth phase: Ideally, freeze cells during the exponential growth phase when they are rapidly dividing. (By doing this you ensure a good yield for future experiments.) • Low passage number: While not a strict rule, freezing cells at a lower passage number (generally between 2-10) is preferred. (It minimizes the risk of mutations or accumulated cellular changes that can occur with repeated passaging.) I hope this helps you!
@qinyiwang4551
5 жыл бұрын
How do you determine the appropriate volume of freezing media?
@thermofisher
5 жыл бұрын
Thank you for your question Qin. The amount of freezing media will be dependent on two factors: the culture vessels (example: Cryo vials) that is being used and the density of the cells. The volume often ranges from 0.5 mL to 4 mL. For additional questions, please contact our Technical Support team at thermofisher.com/askaquestion and we’d be happy to help.
@TheInspiry
4 жыл бұрын
*Can we preserve cells in -70 C for long term storage?*
@thermofisher
4 жыл бұрын
Hi Harry, Thank you for your question. No, cells should be kept in an isopropanol chamber at -80°C for overnight prior to liquid nitrogen storage. For more information about freezing cells, please check the following link. Thank you! www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/freezing-cells.html.
@ArandomStranger-x3r
2 жыл бұрын
You could use a Freeze dryer
@BeBetter26
3 жыл бұрын
Did you use PBS -/- or +/+ for washing cells? Thank you
@thermofisher
3 жыл бұрын
Thank you for your question Nimrah. Can you please reach out to our technical support team so they can reply to your question? They can be reached at thermofisher.com/askaquestion. Thank you!
@kosheeka
3 ай бұрын
I am assuming you the +/+ notation is for the presence of calcium and magnesium ions. For washing purposes calcium and magnesium (+/+) are generally added to the PBS.
@negarshtehrani6380
3 жыл бұрын
What is the freezing media?
@thermofisher
3 жыл бұрын
Thank you for your question. Please contact our technical support team at thermofisher.com/askaquestion for additional information. Thank you!
@동네-e9v
6 жыл бұрын
L929?
@thermofisher
6 жыл бұрын
Yes, our cell freezing medium can be used with L929 cells.
@xperiax4055
6 жыл бұрын
Das sieht sehr interessant aus!
@The15121973
4 жыл бұрын
येस
@NewWesternFront
Жыл бұрын
bonjourno
@BioBat
2 жыл бұрын
what kind of youtube rabbit hole have a stumbled upon bruh
@ch3ckm8
Жыл бұрын
welcome to the nerds world
@sahelidey3146
3 жыл бұрын
Why was the culture centrifuged after trypsinization and the supernatant discarded?
@megasabri173
2 жыл бұрын
you need to change their medium before freezing. If you use DMSO the solution won't form crystals (like water does) and break the cells
@kosheeka
3 ай бұрын
After trypsinization (detaching cells), they spin down the culture in a centrifuge to pellet the cells at the bottom. The supernatant (liquid on top) gets discarded because it contains: • Trypsin: This enzyme detaches cells but can be harmful if left on during freezing. • Dead cells & debris: These won't survive cryopreservation and can harm healthy cells. They want healthy, live cells for freezing, so they keep the cell pellet and wash away the rest!
@Dhanalakshmi-zr9pi
5 жыл бұрын
is this necessary to filter the freezing medium?
@thermofisher
5 жыл бұрын
Thank you for your question. If the freezing media is made in house (ie in your lab), we do recommend filtering the media. However, if you purchase an already made freezing media from us, there is no need to filter. For additional information, please contact our technical support team at thermofisher.com/askaquestion. Thank you!
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