Gulino, Angela Marie.
Insulin secretion dynamics of recombinant hepatic and intestinal cells.
Degree: MS, Biomedical Engineering, 2008, Georgia Tech
Hepatic and intestinal endocrine cells are potentially helpful targets for recombinant insulin expression. As the two cell types exhibit different secretion kinetics,it has been hypothesized that a combination of the two would better approximate insulin secretion kinetics from normal, functioning beta-cells than either cell type alone. This hypothesis was tested using two hepatic cell lines transiently transduced with one of three adenoviruses for insulin expression along with a stably transfected recombinant intestinal L cell line.
The insulin secretion kinetics were analyzed for both the hepatic and intestinal cells to determine the potential of combining them to reproduce the insulin secretion kinetics of a normal, functioning beta-cell. It was observed that the two recombinant hepatic cell lines secreted insulin in a more sustained manner exhibiting slower release kinetics. They also exhibited an increase in insulin secretion when stimulated by the cocktail of nutrient secretagogues (glucose and meat hydrolysate) versus stimulating with only glucose. The cells transduced with the adenovirus containing an additional cytomegalovirus (CMV) promoter and green fluorescent protein (GFP) exhibited the
highest insulin secretion after stimulation, whereas the cells transduced with an adenovirus encoding for destabilized preproinsulin mRNA exhibited the lowest secretion rates.
The recombinant intestinal cell line (GLUTag-INS) secreted insulin with rapid kinetics upon stimulation, apparently due to the presence of secretory granules containing pre-synthesized insulin. The experiments demonstrated that the cells stimulated with medium containing only meat hydrolysate exhibited a significantly higher
insulin secretion relative to secretagogue-free controls. The insulin secretion was not
further enhanced when meat hydrolysate was combined with glucose.
Advisors/Committee Members: Dr. Athanassios Sambanis (Committee Chair), Dr. Barbara Boyan (Committee Member), Dr. Peter Thule (Committee Member).
Subjects/Keywords: Genetic engineering; Diabetes; Liver; Insulin; Kinetics; Intestinal; Liver cells; Intestines; Endocrinology; Recombinant human insulin; Pancreatic beta cells
to Zotero / EndNote / Reference
APA (6th Edition):
Gulino, A. M. (2008). Insulin secretion dynamics of recombinant hepatic and intestinal cells. (Masters Thesis). Georgia Tech. Retrieved from http://hdl.handle.net/1853/28220
Chicago Manual of Style (16th Edition):
Gulino, Angela Marie. “Insulin secretion dynamics of recombinant hepatic and intestinal cells.” 2008. Masters Thesis, Georgia Tech. Accessed January 24, 2021.
MLA Handbook (7th Edition):
Gulino, Angela Marie. “Insulin secretion dynamics of recombinant hepatic and intestinal cells.” 2008. Web. 24 Jan 2021.
Gulino AM. Insulin secretion dynamics of recombinant hepatic and intestinal cells. [Internet] [Masters thesis]. Georgia Tech; 2008. [cited 2021 Jan 24].
Available from: http://hdl.handle.net/1853/28220.
Council of Science Editors:
Gulino AM. Insulin secretion dynamics of recombinant hepatic and intestinal cells. [Masters Thesis]. Georgia Tech; 2008. Available from: http://hdl.handle.net/1853/28220