I want to create a hyperglycemic environment for my beta cells (beta TC cell line -Mus musculus-) as to mimic the environment of a diabetes while comparing it to a normoglycemic control.
Found a set of protocols from previous literature which had different concentrations and I couldn't decide which one to use.
I settled on 5.6 mM as a normoglycemic and 22.0 mM for a hyperglycemic medium,
Would appreciate it if someone suggests better protocol.
Dear Mohammad,
Which cells are you using? Are these beta cell lines or primary beta cells? Beta cell lines grow in different media, e.g., EndoCbH cell lines grow in 5.5 mM glucose, INS1 cells grow in 11 mM glucose and MIN6 cells grow in 25 mM glucose. For primary pancreatic islets, glucose concentrations would vary for a different organism. Though glucose concentration around 5-6 mM is used for normoglycemic conditions in investigation of primary islets and for hyperglycemic conditions, you may find a variety of glucose concentrations (between 8 to 30).
Do you mimic diabetic environment only by changing glucose?
The next question what is your output? In which glucose levels do you think you may see a difference? If it is not known, you may try several concentrations (maybe also go lower than 5.6 mM) at first to identify a dose response or the optimal glucose concentration for your output.
Good luck,
Julia.
Thanks for the quick response Julia,
I should've added all that in the question so apologize for being too amateur as this is still my first research in this field.
I'm using a beta TC cell line from ATCC (mus musculus)
(http://www.lgcstandards-atcc.org/products/all/CRL-11506.aspx?geo_country=ru)
I'm only changing glucose at this stage, and I'm aiming for the activation of a specific signalling pathway (Wnt\B-catenin to be specific)
Previous articles I read have used concentrations varying between 18.5 & 23 mM (For the same output) so I was kind of confused if it would make that big difference in my results.
Thanks again.
Dear Mohammad,
These cells grow normally in 25 mM glucose, therefore anything below that, in my opinion, doesn't represent a good hyperglycemic medium for these cells. Actually, 5.6 mM glucose for these cells would represent starvation.
A better model here is required, as primary islets, INS1 cells, or EndoC cells. Are there any better models than beta TC cell line available for you?
Julia.
Dear Julia,
I checked with a specialized doctor in my university regarding the cell line I have and what other options I can get,
He agreed on what you said regarding the beta TC cell line. However the only other option I have is the 1.1B4 human derived beta cell line, which I read about in literature and found a protocol that used 5.6 mM as normoglycemic & 25 mM as Hyperglycemic environment.
If you have any other information regarding the protocol for this particular cell line I would really appreciate it.
Regards.
Dear Mohammmad,
This cell line grows in lower glucose levels, and therefore 25 mM would be hyperglycemic for it. It may be a good idea for the cells to get used to 5.6 mM before the experiment by incubating the cells in the basal glucose concentration (5.6 mM) for at least O.N. This way all the cells start from the same level of glucose and either stay at the same level or are exposed to hyperglycemia. I hope this helps.
Good luck,
Julia.
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