Russell Mortishire-Smith wrote:
>> I'm a postgraduate student doing a PhD in association with Merck, and
> I am using Biosilon beads from Nunc to immobilize LTK- cells
> for in vitro experiments. The cells grow fine in Dulbecco's medium,
> but when the medium is changed to simple buffer (for the NMR expt)
> containing just NaCl, KCl, MgSO4, phosphate, HEPES &c the cells
> rapidly fall off the beads. I can't use Dulbecco's for the
> experiments.
>> Does anyone have any ideas as to why this should be, or how to get
> around it.
Hi Ailish/Russell,
the 1st thing that springs to mind is a lack of Ca2+ ions in the new
buffer, 1-1.5mM CaCl2 should be enough also is the pH always about 7.4.
I've never worked with Biosilon beads, but some cells don't always like
non plastic surfaces (charge problems) if there is no Calcium or other
free proteins swimming around.
Well this was my experience with RBL-h2 cells, and C6-glioma cells.
yours
Christoph.
BL-Institut für Neurobiologie
Magdeburg
Germany.
> Thanks!
>
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rather famous Rowan man)
