Laminin Use - (Mar/30/2009 )
I have been using a laminin coating to grow some cells. They can also be grown in suspension without the use of laminin. I am now trying to grow them in suspension but despite washing up to 3 times with HBSS at each split, they continue to attach rather than grow in suspension. I expect this is due to residual laminin left on the cells. Is there any way to wash the laminin from the cells more successfully? 
-esklam-
esklam on Mar 30 2009, 05:22 PM said:
I have been using a laminin coating to grow some cells. They can also be grown in suspension without the use of laminin. I am now trying to grow them in suspension but despite washing up to 3 times with HBSS at each split, they continue to attach rather than grow in suspension. I expect this is due to residual laminin left on the cells. Is there any way to wash the laminin from the cells more successfully?
You could try a trypsin-EDTA solution to detach and eliminate the laminin, but this may be pretty hard on the cells. How firmly are the cells attached? I've worked with cells in suspension that weakly adhered to the flask by a sort of electrostatic attraction. There's not much you can do about this, but its not really a problem either, just titurate the media a few times to dislodge. Also, some companies make culture plates designed to enhance attachment, make sure you're not using those.
-gfischer-