attachment HEK-293 cells

Andrew Doherty A.Doherty at Bris.ac.uk
Fri Nov 8 03:49:47 EST 1996


Rob Leurs wrote:
> 
> To study 7TM receptors and to measure their activity by use of cAMP
> measurements we express these receptors in HEK-293 cells. To improve
> attachment of these cells we use Pronectin F attachment factor. Since the
> attachment of these cells is still not very good I want to know if their
> are any other methods to improve the attachment of HEK-293 cells. I would
> also be willing to switch to another cell system with large cAMP responses.
> Any protocols or suggestions for other cells ??
> 
> ******************************************************************************
> Rob Leurs, Ph.D
> Leiden/Amsterdam Center for Drug Research
> Division of Medicinal Chemistry
> Department of Pharmacochemistry, Vrije Universiteit
> De Boelelaan 1083, 1081 HV Amsterdam
> the Netherlands
> 
> fax: 31204447610
> tel: 31204447579
> leurs at chem.vu.nl
Hi Rob,

we use 25uM poly-D-lysine for 5 mins with two washes with water either
side to coat our coverslips to grow HEK293's. We then use them either
for patch clamping, confocal microscopy or immunocytochemistry. They
stick OK with this, but can still come off if treated too roughly. I'm
afraid HEK293's are simply a difficult cell line to get to stick well.
If you need to wash your cells quickly during your experimental
protocol, then I would consider another cell line. CHO's and BHK's are
very tough and do not require any attatchment factors to still REALLY
well, but if you're doing transient expression, I would gice COS cells a
go - I don't think they require anything to make them stick, but don't
quote me on that!!!!

Hope it helps
Andy D
-- 
*************************************************************
Dr Andrew Doherty		email -  a.doherty at bris.ac.uk
Dept. Anatomy			Tel (0117)9287421
School of Medical Sciences	Fax (0117)9287402
University of Bristol
University Walk
Bristol UK
BS8 1TD
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