I'm looking for a culture protocol for MS1 cells (Mile Sven 1, ATCC: CRL-2279 ). This is an adherent endothelial cell line isolated from mouse pancreas. ATCC has a protocol, but it's lacking details on:
1) How often should they be passaged
2) How many cells should be seeded in a T75 flask
Thank you in advance for any help
Pia Nyeng ATCC should have the protocol on their website. They recommend a subculture ratio of 1:4 to 1:8 which should be passaged 2-3 times a week.
In terms of how many cells should be seeded into a T75 flask, ThermoFisher has a detailed table with useful information: https://www.thermofisher.com/uk/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html
Please bear in mind that these numbers are for HeLa cells which are quite large and are cancerous. You may need to adapt the numbers but this should be a good starting point.
Dear Jack Hopkins
Thank you for answering my question.
What I am looking for, is the advice of someone who is working with the cells and have an optimized protocol.
I already have the ACCT protocol, but they do not indicate how often the cells should be passaged (it's change of medium every 2-3 days), and at which density the cells thrive the best.
Pia Nyeng you should maintain cells at between 30-80% confluency. Any less will induce a lag phase as the cells adjust to their surroundings. Any higher will induce a stationary phase as cells run out of resources and space.
My recommendation would be to allow your cells to grow to 80% confluency, and passage at a 1:8 ratio. This gives you three cell cycles for growth (2, 4, 8). If you know how long your cells take to divide, you can multiple this by three and come up with a protocol as to when to passage your cells.
I hope this helps!
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