Gene expression of purified β-cell tissue obtained from human pancreas with laser capture microdissection

Lorella Marselli, Jeffrey Thorne, Yu Bae Ahn, Abdulkadir Omer, Dennis C. Sgroi, Towia Libermann, Hasan H. Otu, Arun Sharma, Susan Bonner-Weir, Gordon C. Weir

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63 Scopus citations

Abstract

Context: Human β-cell gene profiling is a powerful tool for understanding β-cell biology in normal and pathological conditions. Assessment is complicated when isolated islets are studied because of contamination by non-β-cells and the trauma of the isolation procedure. Objective: The objective was to use laser capture microdissection (LCM) of human β-cells from pancreases of cadaver donors and compare their gene expression with that of handpicked isolated islets. Design: Endogenous autofluorescence of β-cells facilitated procurement of purified β-cell tissue from frozen pancreatic sections with LCM. Gene expression profiles of three microdissected β-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 β-cell rich tissue in human pancreas. When compared with array profiles of purified β-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 β-cell enriched tissue from human pancreas sections without the trauma and ischemia of islet isolation.

Original languageEnglish (US)
Pages (from-to)1046-1053
Number of pages8
JournalJournal of Clinical Endocrinology and Metabolism
Volume93
Issue number3
DOIs
StatePublished - Mar 2008

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ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

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). Gene expression of purified β-cell tissue obtained from human pancreas with laser capture microdissection. Journal of Clinical Endocrinology and Metabolism, 93(3), 1046-1053. https://doi.org/10.1210/jc.2007-0931