Hi there, i'm starting an iPSC culture.
My supervisor told me that in their lab it's usually 1:10 ratio split for an 80% confluence iPSC. We're growing the iPSC culture in a 6-well non-coated plate, each of them at 2ml.
My question might be a bit silly, but I want to just confirm that 1:10 split ratio means that I passaged 1 well to 10 well of a new plate, right?
I'm a bit confused because I'm told to grow it only in 6 well (1 plate only) but each of the well has 1/10 of the concentration. It might be just a language barrier or so, but I'd appreciate all the help!
PS. We're adding the cells as an aggregate, so we do not count them.
Hello Juan Dennis Bahrian
For a 1:10 split ratio, you may take 200ul of cell suspension of 80% confluent iPSC culture and add 1800ul of fresh culture media to each well of a 6-well plate so that each well will contain a total of 2ml (cells + media).
Best.
So yes, you are correct that for a 1:10 split ratio, you would typically split the contents of **one well into 10 wells** of a new plate. However, if you're only using **1 plate with 6 wells**, it may mean that instead of transferring to 10 wells, you’re just diluting the cells in the existing wells to **1/10th of the concentration**.
For example:
1. You would collect the cells from 1 well.
2. You can then dilute them across the 6 wells on the same plate.
3. This dilution would lead to a reduced cell density in each well, equivalent to splitting at a 1:10 ratio (even though you're not using exactly 10 wells, the concentration is still lowered).
Since you're working with aggregates and not counting cells, it's more about ensuring the relative concentration of cells in the new wells is about 1/10th of the original.
It’s always good to check with your supervisor if they mean a literal 1:10 split or just a significant dilution while staying within one 6-well plate!
- Badges
- Science topic
- Similar topics
- Engineering
- Materials Engineering
- Powders