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The following message was posted to: PharmPK
Dear All,
I am trying to establish accelerated 7-day caco2 culture for which I =20
varied parameters like seeding density and filter type (polycarbonate =20=
and polyester).
The polyester had certain advantages over polycarbonate:
* It allowed cell growth visibility
* Had higher TEER value
* Low % Lucifer yellow permeability (~ 0.5%)
Therefore for further standardization polyester filters were used. =20
However the Papp of Caffeine determined was very low. To blame higher =20=
tight junction will be unfair as the Lucifer yellow permeability was =20
between acceptable ranges.
Therefore my concern now is about the compound retention to the =20
filter. But I can=92t ignore the fact that caffeine being a hydrophilic =20=
drug has negligible chance of binding to the filter.
Recovery attained for caffeine in the experiment was around 60% after =20=
establishing a good linearity (RSQ=3D0.99) with transport buffer.
Can anyone help me with any solution to my problem
I will be looking forward to expert opinions
New Chemical Entity Research
Lupin Research Park
46A/47A, Village: Nande, Taluka: Mulshi
Pune-411 042, India=
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The following message was posted to: PharmPK
Dear All,
Thanks for all of your valuable inputs due to which I was able to
establish the traditional 21-day Caco2 culture and also the
accelerated 7-10 day Caco2 culture.
In my previous mail I had mentioned about low Papp of Caffeine and my
concern over its non-specific binding to membranes. However this issue
has been sorted in latter experiments where recovery is more than 85%.
I am using Caco2 subclone instead of parent Caco2 cell line, perhaps
this might be the reason for low Papp of Caffeine as subclones are
more homogeneous in nature and this may contribute to tighter junctions.
Hope this finding is relevant and expecting any suggestions for
further refinement.
Regards,
Pankajini
New Chemical Entity Research
Lupin Research Park
46A/47A, Village: Nande, Taluka: Mulshi
Pune-411 042, India
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