Working with beta cells

[jaykay]

Hi all

I work with INS1E cells, a beta cell line, for insulin secretion studies. My experiment is to give each well of cells an increasing amount of glucose, and in theory the beta cells should release increasing amounts of insulin. However my results have been inconsistent even with careful attention to every step. Does anyone have any thoughts on why this problem occurs?

[DrDNA]

What is your positive control or is this supposed to be your positive control?

Where did you get the cells and how were they characterized?

[jaykay]

Well we are using glucose plus EGCG (green tea compound) which is supposed to stimulate insulin release. So we have glucose at say, 3, 5, 11, 16, and 20 mM. Then we add EGCG or do not add it to each glucose concentration. I don't know where my PI got the beta cells from, some acquaintence at a university I think. What do you mean by how were they characterized.

[DrDNA]

Well we are using glucose plus EGCG (green tea compound) which is supposed to stimulate insulin release. So we have glucose at say, 3, 5, 11, 16, and 20 mM. Then we add EGCG or do not add it to each glucose concentration. I don't know where my PI got the beta cells from, some acquaintence at a university I think. What do you mean by how were they characterized.

 

I think you are saying that are you are getting the same responce with and without the EGCG. Is that correct?

Do you have something else to stimulate a similar response? If so, have you tried it? If not, can you? (positive control)

 

Did these particular cells ever behave as anticipated in your hands or in someone in your lab's hands?

Where did you get the EGCG? Does YOUR prep (the very same one that you are using) work as anticipated with someone else's cells?

How are you measuring the insulin? Do you have positive controls for the instrument?

Finally, something like this can often be traced back to a simple decimal point type error......at least when I do the math....