I am planning to analyze the glucose uptake in 3T3-L1 cells. The suggested treatment scheme will be used to analyse the effect of plant extract on glucose uptake. Control groups (treated/untreated): glucose+extract, glucose, extract.
How long should the period of treatment with glucose be?
Do you suggest some different model for glucose uptake and insulin-like activity analysis?
Hi Oskan,
I do not work in the field of glucose transport but Caco-2 cells may be a model. See this paper:
http://www.ncbi.nlm.nih.gov/pubmed/21943636
Best regards
Victor
Good morning Oskan,
In my opinion, the treatments are correct if you want to study the effect on glucose uptake.
In relation to the first question, you can take other studies as a reference. In this paper, http://www.sciencedirect.com/science/article/pii/S0021915001007195#, human aortic endothelial cells were exposed to glucose for 24 hours. However, you can make a pre-assay at different times (12, 24, 48,...) to test if other time-treatment is better for your experiment.
In relation to the second question, I have no experience in up-take of glucose. However, I worked with phenolic compounds. I treated my cells with urolithins and their glucuronides, and after treatment the concentration in the culture medium was analyzed by HPLC (http://www.ncbi.nlm.nih.gov/pubmed/22648625). Furthermore, other authors investigated the presence of metabolites both culture medium and cells lysates (http://www.ncbi.nlm.nih.gov/pubmed/19469472).
Faria et al. (2011) (http://pubs.rsc.org/en/content/articlelanding/2011/FO/C0FO00100G#!divAbstract) describe some transporters implied in the transport of glucose, in blood brain barrier, which could be a good markers to investigate the effect of glucose on your cells.
If you need something or you have any doubt, do not hesitate to ask me. If you have no access to the articles I have described above and you need it, please let me know and I send you them
All the best,
Juan.
The important question here is - would propagation/ differentiation of 3T3-L1 in high-glucose (4.5g/L) DMEM would make the cells diabetic? It's something we're currently discussing about at our lab
Yes I want to look on glucose uptake!
I wonder about the culture media, that should be used. As you said, I should find the best glucose concentration and the treatment time.
Thanks to bouth of you! I will read that papers and look for more Juan! Petar you know we always discuss the 3T3-L1 metabolism, your observation are usefull! Good point thank you!
Hi Oskam, im also doing glucose uptake for my plant extract, plz, could tell me the best excitation and emission. in addition, if u have any assay or any info. regarding glucose up take for diabetic cells
Best regards
I am using the Glucose uptake kit from Abcam (colorimetric, which is more sensitive to the fluorimetric one!). Not cheap, but works nice!
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