What kind of speed for HepG2 cells. - (Sep/22/2009 )

Hi, I get confused about spin speed of HepG2 cells. I found some dead cells in dishes. Then i use low speed to discard these deads. It is 500g. Actually i dont effectively elimate dead cells. however i lost a lot of cells.
The problems,
1. what speed do you use for HepG2. and this speed will not harm the cells. please tell me the speed as G, not rpm.
2. it is always some dead cells attched on the surface of alive cells. how do i deal with it.

Thanks a lot to all of you.

shawnpeter on Sep 23 2009, 03:42 AM said:

WOW......500g is far too fast for just pelleting cells.

1. I use about 100g on the ALC centrifuges that I use for TC work. The speed is very important in terms of cell viabilty... the greater the speed , the greater the cell damage.

2. Trypsinisation will also cause a loss of some cells. In a population of cells (after trypsinisation) there will be a mixture of:
Dead cellls
Viable cells
Cells which may senence
Cells which will eventually apoptose.

You can reduce the number of dead cells attached to live cells by:
Reducing the speed of spin
Reducing the time the trypsin is in contact with the cells
Let the cells attach to the flask for 30 minutes, then wash away the cells that are still non adherent...you will lose some viable cells, but the dead cells will be washed away

Hope this is useful

Kindset regards

Rhombus

Let the cells attach to the flask for 30 minutes, then wash away the cells that are still non adherent...you will lose some viable cells, but the dead cells will be washed away

thanks for your reply.
i dont understand this words. Just without treatment, let cells sit in the hood, is that right?