4 replies to this topic

[bcsein]

Hi,
I am going to start MCF-7 cultures and need some advice about:
use RPMI or MEM with glutamine?
use medium without phenol red for treatments in all cases? How to do this, how long before treatment without phenol red?
Which way to trysine ? These cells are doing junctions very fast and it is diffciult to disgragate them.
I really would appreciate your advice.
Thanks

[bob1]

Use the medium they have last been cultured in, preferably with L-glutamine and insulin, but without phenol red as this will slow the growth rate.  Trypsinise as you would other cells, you may need to add the trypsin for longer and be a bit more vigorous with the pipetting to make a single cell suspension.

[bcsein]

bob1, on Oct 27 2009, 03:40 PM, said:

Use the medium they have last been cultured in, preferably with L-glutamine and insulin, but without phenol red as this will slow the growth rate.  Trypsinise as you would other cells, you may need to add the trypsin for longer and be a bit more vigorous with the pipetting to make a single cell suspension.

Thanks a lot. The medium they have now is with phenol red, should I change them ? They seem to be growing well although I can see big  vauoles in the culture.

[bob1]

They will grow in phenol red, but it does inhibit the growth and make them more prone to dying.  I would change them to phenol red free medium.

[bcsein]

bob1, on Oct 28 2009, 04:16 PM, said:

They will grow in phenol red, but it does inhibit the growth and make them more prone to dying.  I would change them to phenol red free medium.

Thank you very much