splitting cellsideas?
Moderators: honeev, Leonid, amiradm, BioTeam
splitting cellsideas?
My protocol says :
Hela cells divide once every 24 hours. The split ratio was thus (1/2)^x where x is the number of days to the next cellculture procedure. When the day comes detach the cells and then add 8ml MEM++ and from the cell suspension, (1/2)^x *8ml was transfered into a new culture flask and MEM++ was added to a total volume of at least 25ml.
My questions:
1. Why is the split ratio (1/2)^x when the cells divide once every 24 hours?
2. What does it mean with (1/2)^x *8ml? Is that the cell number or the volume?
Hope very much for ideas.
Thanks.
Hela cells divide once every 24 hours. The split ratio was thus (1/2)^x where x is the number of days to the next cellculture procedure. When the day comes detach the cells and then add 8ml MEM++ and from the cell suspension, (1/2)^x *8ml was transfered into a new culture flask and MEM++ was added to a total volume of at least 25ml.
My questions:
1. Why is the split ratio (1/2)^x when the cells divide once every 24 hours?
2. What does it mean with (1/2)^x *8ml? Is that the cell number or the volume?
Hope very much for ideas.
Thanks.

 Garter
 Posts: 1
 Joined: Fri Jun 30, 2006 12:47 pm
Splitting cells
1) if your cells are confluent in flask/plate with the same area as the flask you want to perform an experiment on your cells. Supposing you split them in half: as they double every 24h (i.e. the area they cover doubles) they will be confluent again in one day.
If you split them 1/4, they will be confluent in 2 days (they will have to double twice 1/2^2=1/4).
if you split them 1/8 they will be ready in 3 days, they will have to double 3 times (1/2^3=1/8). 4 days: 4 doublings, (1/2^4=1/16)
2) i assume that the 8 ml is the total volume that you will seed your plate or flask with. I would ignore this, it makes it overcomplicated. just divide your cells as per the formula above. Think about the area you want to cover with your cells, this is the important part to consider.
If you split them 1/4, they will be confluent in 2 days (they will have to double twice 1/2^2=1/4).
if you split them 1/8 they will be ready in 3 days, they will have to double 3 times (1/2^3=1/8). 4 days: 4 doublings, (1/2^4=1/16)
2) i assume that the 8 ml is the total volume that you will seed your plate or flask with. I would ignore this, it makes it overcomplicated. just divide your cells as per the formula above. Think about the area you want to cover with your cells, this is the important part to consider.
Who is online
Users browsing this forum: No registered users and 2 guests