I want to determine the drug effect in inhibiting human bronchial smooth muscle contraction induced with histamine. Currently, I tried it with
1. 0.5 X 10^6 cell per well in 24 well plate with 2 mg/ml collagen gel
- histamine concentration : 10 - 100 uM
2. 0.2 X 10^6 cell per well in 24 well plate with 1.5 mg/ml collagen gel
- histamine concentration : 10 - 100 uM
Both conditions did not work. The induced group showed no difference with the normal control group.
And, for the 2 mg/ml collagen gel, the spindle shape of the cell did not look normal as compared to the 1.5 mg/ml collagen gel.
Any recommendations to improve my assay?
Thanks.
Best regards,
Hui Min
(PhD student)
I would look at the paper from Judith Black's group who have used this technique successfully many time, in this study to compare ASM cells from healthy and asthmatic patients. The source of the ASM cells should not be important, but the culture conditions are extremely important.
http://thorax.bmj.com/content/62/10/848.long
Hi Chris, I measured the surface area of the gel at 0, 15, 30, 45 and 60 minuste after addition of histamine. Thanks. :)
Hi, Jeremy, ok, i will refer to that paper and follow exactly the same protocol try again. Thanks. :)
Hello Huimin,
I also had the same problem. I used acetylcholine to stimulate the cell but did not observe any contraction.
Have you found the solution? If yes, would you mind sharing with me?
Thank you very much,
Duy
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