An optimized in vitro assay for screening compounds that stimulate liver cell glucose utilization with low cytotoxicity.
BACKGROUND
Screening compounds that stimulate liver cell glucose utilization with low cytotoxicity plays an important role in anti-diabetic drug discovery. Existing in vitro assay for the quantitative measurement of cell glucose utilization rate and cytotoxicity of compounds needs to be modified in order to improve the simplicity and reproducibility. An optimized assay was described to addresses these problems.


METHODS
Compounds were directly mixed with cell suspension. After 20 h incubation, glucose decrement detection in culture medium was performed, followed by the assessment of cell viability using Cell Counting Kit-8 (CCK-8).


RESULTS
A series of experiments were conducted to define optimal conditions such as cell inoculation density, incubation time and CCK-8 concentration in cell culture medium. Glucose utilization unit (GUU) was defined for the evaluation of both efficacy and cytotoxicity of a compound. It was found that Rosiglitazone significantly stimulated cell glucose utilization with negligible cytotoxicity at the concentration of 10(-5) M.


CONCLUSIONS
The simplicity and efficiency of the optimized method has been proven in this study. Our findings show that this assay has potential application in anti-diabetic drug discovery such as new peroxisome proliferator-activated receptor gamma (PPARgamma) agonists.
Zou
Chenhui
C
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xiannongtan Street, Beijing 100050, China.
Shen
Zhufang
Z
D002227:Carbazoles / Q000494:pharmacology; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005947:Glucose / Q000378:metabolism*; D005905:Glyburide / Q000494:pharmacology; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology* / Q000633:toxicity; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008687:Metformin / Q000494:pharmacology; D011422:Propionates / Q000494:pharmacology; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000494:pharmacology
D002227:Carbazoles; D007004:Hypoglycemic Agents; D007328:Insulin; D011422:Propionates; D045162:Thiazolidinediones; C515629:chiglitazar; D000077154:Rosiglitazone; D008687:Metformin; D005947:Glucose; D005905:Glyburide