Author/Authors :
Mohammadi Farsani, Taiebeh Department of Medical Biotechnology - School of Advanced Technologies in Medicine - Tehran University of Medical Sciences - Tehran, Iran , Motevaseli, Elahe School of Advanced Technologies in Medicine - Tehran University of Medical Sciences - Tehran, Iran , Neyazi, Nadia Department of Medical Biotechnology - School of Advanced Technologies in Medicine - Tehran University of Medical Sciences - International Campus (TUMS- IC)- Tehran, Iran , Khorramizadeh, Mohammad Reza Biosensor Research Center - Endocrinology and Metabolism Molecular-Cellular Sciences Institute - Tehran University of Medical Sciences - Tehran, Iran , Zafarvahedian, Elaheh Department of Pharmacology and Toxicology - Faculty of pharmacy - Tehran University of Medical Sciences - Tehran, Iran , Ghahremani, Mohammad Hossein Department of Pharmacology and Toxicology - Faculty of pharmacy - Tehran University of Medical Sciences - Tehran, Iran
Abstract :
Insulin-degrading enzyme (IDE) is a conserved zinc metallopeptidase. Here, we have evaluated the
effect of passage number and culture time on IDE expression and activity in colorectal adenocarcinoma cell line
(Caco-2). Methods: Caco-2 cells were cultured with different passage ranges of 5-15, 25-35, 52-63 for 48, 72, and
120 hours. Subsequently, IDE expression and enzyme activity were assessed by Western blot analysis and
fluorescent assay, respectively. Results: Our results confirmed that the amount of IDE was higher in cell extract
compared to supernatant, and different passage numbers and culture times had small effect on IDE expression.
However, when cells were cultured in the passage number range of 5-15 for 72 hours, the IDE activity was
35% higher compared to other passage numbers (p < 0.05). Conclusion: The use of Caco-2 cells at passage
number range of 5-15 and culture time of 72 hours provides proper conditions for IDE-related studies.
