Protocol Online logo
Top : Forum Archives: : Cell Biology

Prostate cancer PC3 cells - Doesn't spread on glass well, doesn't migrate well, arghhh (Jul/05/2008 )

Hi all,
Was hoping that there'll be someone experienced with PC3 cells.
I'm trying to measure migration rates in stable cell lines of PC3.

In a woundhealing (scratch assay) model using video microscopy first, they don't really spread on the glass, then they didn't migrate at all (moved on the spot) for 16 hours.

In a transwell assay (0.8 um pore) which I used successfully for fibroblasts, I didn't get ANY cells migrating toward serum.

I think I need to coat the glass and the transwell filter with something. Any experience/suggestion? I have collagen or fibronectin in the lab, but can also buy in others if they are known to work.

-Michelle4-

QUOTE (Michelle4 @ Jul 5 2008, 04:49 AM)
Hi all,
Was hoping that there'll be someone experienced with PC3 cells.
I'm trying to measure migration rates in stable cell lines of PC3.

In a woundhealing (scratch assay) model using video microscopy first, they don't really spread on the glass, then they didn't migrate at all (moved on the spot) for 16 hours.

In a transwell assay (0.8 um pore) which I used successfully for fibroblasts, I didn't get ANY cells migrating toward serum.

I think I need to coat the glass and the transwell filter with something. Any experience/suggestion? I have collagen or fibronectin in the lab, but can also buy in others if they are known to work.


try to coat glass (poly-L-lysine, fibronectin, collagen etc)

pore size seems to be very tight; offer larger pores (3 ┬Ám or more)

-The Bearer-

Oops, I meant 8 um pores.
Thanks for the suggestions smile.gif I was hoping someone would have tried different ECM coats so that I don't need to try all of them.

-Michelle4-

QUOTE (Michelle4 @ Jul 5 2008, 03:36 PM)
Oops, I meant 8 um pores.
Thanks for the suggestions smile.gif I was hoping someone would have tried different ECM coats so that I don't need to try all of them.

Let me make it up to you sleep.gif

Did you coat with Matrigel?

..

-cellcounter-