5 replies to this topic

[swarna]

i would like to perform toxicity assays (cell titre glo assay and toxilight bioassay) on HL 60 cell line
before i start i would like to know when can i add my test compunds to the culture?
and how many cells can i use per ml

is there any protocols to follow before i start these Assays?

Edited by swarna, 25 February 2009 - 03:07 AM.

[bob1]

When you add your compounds depends on the aim of your experiment, typically you should add your compounds after the cells have adhered and recovered from seeding (24-48 hours post seeding).  However, if you are doing something like transfecting and then drug treating, you will need to determine when the transfection has worked and then add your drugs.

The number of cells per ml will depend on the assay: read the instruction manual.

[swarna]

bob1, on Feb 25 2009, 04:52 PM, said:

When you add your compounds depends on the aim of your experiment, typically you should add your compounds after the cells have adhered and recovered from seeding (24-48 hours post seeding).  However, if you are doing something like transfecting and then drug treating, you will need to determine when the transfection has worked and then add your drugs.

The number of cells per ml will depend on the assay: read the instruction manual.

the probelem is these are suspension cells, i sorted out the number of cells i need to use
but i still confused when to add the test compound (iam doing this assay to find at concentrations the cells start to die)

Edited by swarna, 26 February 2009 - 02:14 AM.

[Minnie Mouse]

I think you can treat HL60 cells immediately after seeding.
They don't need to recover from trypsinize nor waiting them to adhere to the bottom of the plate/flask.
As long as you don't spin them too hard before seeding them.

Hope this may help.

[bob1]

As HL60 is suspension, they should be fine to treat any time.  

It seems you will have to do a titration of dose and time (i.e. set up several different concentrations and harvest each concentration at several time points) to see the effects of your drugs, weaker concentrations may still work, it may just take longer than high concentrations.

[swarna]

bob1, on Feb 26 2009, 04:30 PM, said:

As HL60 is suspension, they should be fine to treat any time.  

It seems you will have to do a titration of dose and time (i.e. set up several different concentrations and harvest each concentration at several time points) to see the effects of your drugs, weaker concentrations may still work, it may just take longer than high concentrations.

Thank you bob
i will try like that ...i hope it works