thawed cells wont adhere - pc3 and LNcaP (Sep/08/2010 )

The cells are old (1992) and I dont know who exactly froze these cells. I was still in high school but anyway, I thawed them as I usually do any other cells. They are VIABLE they just wont adhere to the flask. Its strange that both cells lines are behaving this way. I have given them extra serum and changed the flask and dishes to see if that might get them to adhere but alas, they are all floating happily. I have never worked with prostate cells before so I am not sure what is going on here. Anyone???

Firstly:
1. are you using tissue culture treated flasks?

2. are you SURE they are viable? How have you decided/checked this?

3. are you using the right media formulation for each cell type?

Now to look at each cell line:
PC3=
if you look at the ATCC info for your PC3 line here (http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=CRL-1435&Template=cellBiology) you will see that these cells can be adapted for suspension culture- is there anyone in your lab you can check with to see if this has been done for your batch? (also worth checking how this is done, perhaps your culture conditions are contributing?)

LNcaP=
again from ATCC (http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=CRL-1740&Template=cellBiology). These cells don't adhere very well and so you shouldn't do anything to them 48hours after thawing to allow them to settle and start to adhere

Hope this helps

Dear Member,

For cells that do not adhere to normal TC plastic, you can use:

Laminin.
Collagen.
Fibronection.
Poly-D-Lysine.
Primaria (special TC plastic from BD).

Kindest regards.

Uncle Rhombus.