Help: cell adhesion assay

David Winterbourne sghk100 at sghms.ac.uk
Fri Sep 4 05:15:39 EST 1998


Zbigniew Rudzki wrote:
> 
> Oh yes, but fixing the cells you can also attach the nonadherent cells to
> your substrate.  For sure this can occur when the crosslinkers like
> formaldehyde are used.

As the density of the fixative is greater than the cells, cells that have
not settled are prevented from contacting the surface of the well. When I
was setting up this test, I observed that such cells were removed during
the washing steps - I also saw that dead cells were removed.

I agree that cells that have made some initial contacts (attachments?)
with the surface may well have these stabilised. Cells that have attached
but not spread due to the absence of the required factors were fixed to
the surface. However, I cannot say what degree of attachment is sufficient
to overcome this flotation effect or the subsequent shear forces during
the washing steps.

The process of cell attachment in vitro is clearly quite complex: part of
the initial rate is presumably due to settling of the cells from the
suspension seeded in the well. This will depend in part on the depth of
the medium and its viscosity and is presumably of little biological
interest.

-- 
David Winterbourne
Department of Surgery
St. George's Hospital Medical School, London SW17 0RE, England
Tel: 0181-725-5581   Fax: 0181-725-3594



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