31 Marselli, L. Thorne, J. Ahn, Y. B. Omer, A. Sgroi, D. C. Libermann, T. Otu, H. H. Sharma, A. Bonner-Weir, S. Weir, G. C. 2008 J Clin Endocrinol Metab 93 3 1046-53 Mar 0021-972X (Print) 18073315 Lasers Insulin-Secreting Cells/*metabolism Humans *Gene Expression Profiling RNA/analysis Oxidative Stress Oligonucleotide Array Sequence Analysis Mitogen-Activated Protein Kinases/genetics Microdissection/*methods CONTEXT: Human beta-cell gene profiling is a powerful tool for understanding beta-cell biology in normal and pathological conditions. Assessment is complicated when isolated islets are studied because of contamination by non-beta-cells and the trauma of the isolation procedure. OBJECTIVE: The objective was to use laser capture microdissection (LCM) of human beta-cells from pancreases of cadaver donors and compare their gene expression with that of handpicked isolated islets. DESIGN: Endogenous autofluorescence of beta-cells facilitated procurement of purified beta-cell tissue from frozen pancreatic sections with LCM. Gene expression profiles of three microdissected beta-cell samples and three isolated islet preparations were obtained. The array data were normalized using DNA-Chip Analyzer software (Harvard School of Public Health, Boston, MA), and the lower confidence bound evaluated differentially expressed genes. Real-time PCR was performed on selected acinar genes and on the duct cell markers, carbonic anhydrase II and keratin 19. RESULTS: Endogenous autofluorescence facilitates the microdissection of beta-cell rich tissue in human pancreas. When compared with array profiles of purified beta-cell tissue, with lower confidence bound set at 1.2, there were 4560 genes up-regulated and 1226 genes down-regulated in the isolated islets. Among the genes up-regulated in isolated islets were pancreatic acinar and duct genes, chemokine genes, and genes associated with hypoxia, apoptosis, and stress. Quantitative RT-PCR confirmed the differential expression of acinar gene transcripts and the duct marker carbonic anhydrase II in isolated islets. CONCLUSION: LCM makes it possible to obtain beta-cell enriched tissue from human pancreas sections without the trauma and ischemia of islet isolation. DK36836/DK/NIDDK NIH HHS/United StatesU19DK6125/DK/NIDDK NIH HHS/United StatesU4Z RR 16606/RR/NCRR NIH HHS/United StatesJournal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov'tUnited States http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18073315