F.W.J. Boot (Leiden)
26 november 2020 bij 17:02- Antwoorden
Hello Warner, we were wondering how cell viability was measured (TUNEL staining, FDA/PI, MTT)? Thank you!
F.W.J. Boot (Leiden)
26 november 2020 bij 17:05- Antwoorden
(since you also display >100% viability for the TNF-alpha, how are these calculations done, how would you explain this, is there cell death in your control?)
C.W. Hoornenborg (Groningen)
26 november 2020 bij 17:29- Antwoorden
Time has some effect on cell viability, so there could be some cell death in our control but this is also seen in other studies (and with other cell lines). What we think is that the TNF-alpha and ST could also have a proliferation effect. We are checking this as well however, we are waiting for the results.
C.W. Hoornenborg (Groningen)
26 november 2020 bij 17:05- Antwoorden
Hello Fransje, Thank you for your question.
We used the MTT assay for the cell viability measurements.
Best,
Warner
Marleen van Greevenbroek
26 november 2020 bij 18:17- Antwoorden
thank you for this interesting presentation. there seems to be a narrow ‘window of opportinity’ for this compound (i.e. functional concntration). How do you propose to go forward with these experiments?
C.W. Hoornenborg (Groningen)
26 november 2020 bij 18:41- Antwoorden
Marleen, thank you for your question.
Indeed it looks like a narrow ‘window of opportunity’. This was partly because of practical problems with the dosage of ST. It was difficult to increase the dose of ST because the volume of all our test compound would be higher than 20% of the total volume of the wellsif we went higher with our ST dose. Going lower than 80% with the medium resulted in a decrease in cell viability so that would give a confounding effect
But of importance was that we saw an effect in vitro (as we already have shown an effect in vivo see below). And we are now focussing on the mechanism of the effect of ST, so we are looking at the caspase pathway and we are looking at the bioactive compounds present in ST
Also of interest: Our lab also performed experiments in a type 2 diabetic rat model, where 200 mg/mg of ST was used. In this study blood glucose levels were decreased and beta-cell mass was increased after treatment with ST.
Again thank you for your question, and I hope this answer gives some clarity
6 Reacties
Hello Warner, we were wondering how cell viability was measured (TUNEL staining, FDA/PI, MTT)? Thank you!
(since you also display >100% viability for the TNF-alpha, how are these calculations done, how would you explain this, is there cell death in your control?)
Time has some effect on cell viability, so there could be some cell death in our control but this is also seen in other studies (and with other cell lines). What we think is that the TNF-alpha and ST could also have a proliferation effect. We are checking this as well however, we are waiting for the results.
Hello Fransje, Thank you for your question.
We used the MTT assay for the cell viability measurements.
Best,
Warner
thank you for this interesting presentation. there seems to be a narrow ‘window of opportinity’ for this compound (i.e. functional concntration). How do you propose to go forward with these experiments?
Marleen, thank you for your question.
Indeed it looks like a narrow ‘window of opportunity’. This was partly because of practical problems with the dosage of ST. It was difficult to increase the dose of ST because the volume of all our test compound would be higher than 20% of the total volume of the wellsif we went higher with our ST dose. Going lower than 80% with the medium resulted in a decrease in cell viability so that would give a confounding effect
But of importance was that we saw an effect in vitro (as we already have shown an effect in vivo see below). And we are now focussing on the mechanism of the effect of ST, so we are looking at the caspase pathway and we are looking at the bioactive compounds present in ST
Also of interest: Our lab also performed experiments in a type 2 diabetic rat model, where 200 mg/mg of ST was used. In this study blood glucose levels were decreased and beta-cell mass was increased after treatment with ST.
Again thank you for your question, and I hope this answer gives some clarity