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Logo of bmcbiotBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Biotechnology
 
BMC Biotechnol. 2012; 12: 64.
Published online Sep 19, 2012. doi:  10.1186/1472-6750-12-64
PMCID: PMC3469342
Evaluation of insulin expression and secretion in genetically engineered gut K and L-cells
Zalinah Ahmad,corresponding author#1,2 Mina Rasouli,#1 Ahmad Zaid Fattah Azman,3 and Abdul Rahman Omar1
1Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, Universiti Putra Malaysia, UPM Serdang, Selangor, 43400, Malaysia
2Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, UPM Serdang, Selangor, 43400, Malaysia
3Department of Community Health, Faculty of Medicine & Health Sciences, Universiti Putra Malaysia UPM Serdang, Selangor, 43400, Malaysia
corresponding authorCorresponding author.
#Contributed equally.
Zalinah Ahmad: zalinah/at/medic.upm.edu.my; Mina Rasouli: rasouli.mina/at/gmail.com; Ahmad Zaid Fattah Azman: zaid/at/medic.upm.edu.my; Abdul Rahman Omar: aro/at/ibs.upm.edu.my
Received May 17, 2012; Accepted September 17, 2012.
Abstract
Background
Gene therapy could provide an effective treatment of diabetes. Previous studies have investigated the potential for several cell and tissue types to produce mature and active insulin. Gut K and L-cells could be potential candidate hosts for gene therapy because of their special features.
Results
In this study, we isolated gut K and L-cells to compare the potential of both cell types to produce insulin when exposed to similar conditions. The isolated pure K and L-cells were transfected with recombinant plasmids encoding insulin and with specific promoters for K or L-cells. Insulin expression was studied in response to glucose or meat hydrolysate. We found that glucose and meat hydrolysate efficiently induced insulin secretion from K and L-cells. However, the effects of meat hydrolysate on insulin secretion were more potent in both cells compared with glucose. Results of enzyme-linked immunosorbent assays showed that L-cells secreted more insulin compared with K-cells regardless of the stimulator, although this difference was not statistically significant.
Conclusion
The responses of K and L-cells to stimulation with glucose or meat hydrolysate were generally comparable. Therefore, both K and L-cells show similar potential to be used as surrogate cells for insulin gene expression in vitro. The potential use of these cells for diabetic gene therapy warrants further investigation.
Keywords: Diabetes gene therapy, Insulin expression, K-cells, L-cells
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