Serious help in cell culture-Dynabeads

[EndothelialCell]

I am absolutely desperate right now! I am trying to culture cells from blood from patients with scleroderma but the cells keep dying!

This is my first research position and I have gone through 2 months and the cells keep dying. I am able to isolate the cells fine, but I can't culture them.

I am using dynabeads in the procedure. I haven't been heating the medium before I use it, however, I found out earlier this week that I was supposed to. Do you think this could be the reason they are dying?

Also, how do you change medium for cells?

What other techniques can you use??

[Lise]

Hi there

I'm with the Invitrogen Dynal Technical Support, could I ask what Dynabeads product you are using? Normally endothelial cells will grow fine in culture, even with the beads attached. What ratio of beads:cells have you been using?

Your medium could be the problem, the best would probably be to get in contact with the manufacturer of the medium. Did you try to culture the cells after you heated the it?

In order to change the medium you spin down your cells. However, since endothelial cells are adherent you should be able to pipette off the medium without spinning. You will of course loose all cells not adhered to the plastic of the wells/flasks.

Best,
Lise

[EndothelialCell]

Hi Lise!
I was using Dynabeads CD31 Endothelial cell coated with some antibody. What type of Dynabeads do you suggest are best for circulating endothelial cells isolated from sick patients blood??

Thanks!

[wr_1a]

thanks