Publication

Effective removal of alginate-poly-L-lysine microcapsules from pancreatic islets by use of trypsin-EDTA

de Groot, M., Leuvenink, HGD., Fekken, S., Schuurs, TA., van Schilfgaarde, R. & KEIZER, J., 1-Nov-2003, In : Journal of Biomedical Materials Research. Part A. 67A, 2, p. 679-683 5 p.

Research output: Contribution to journalArticleAcademicpeer-review

  • M de Groot
  • HGD Leuvenink
  • S Fekken
  • TA Schuurs
  • R van Schilfgaarde
  • J KEIZER

Although the transplantation of alginate-poly-L-lysine-alginate encapsulated islets of Langerhans usually is successful, graft survival is still limited. Molecular analysis by RT-PCR of the encapsulated islets may provide insight into the mechanisms that affect islets during graft failure. However, RT-PCR on encapsulated islets is not possible because the poly-L-lysine of the capsule interferes with both cDNA synthesis and PCR amplification. We applied a method that mechanically removes the microcapsules from the islets after a short trypsin-EDTA treatment (decapsulation), thereby enabling RT-PCR analysis. The results of this study show that the decapsulation procedure does not affect islet vitality and has only minor effects on islet function and morphology. The decapsulation does not affect GAPDH, beta-actin, Bcl-2, or Bax gene expression. This method is an improvement over the time-consuming manual dissection of microcapsules because it allows for the rapid and relatively harmless removal of capsules on a larger scale. Decapsulation offers the possibility of applying RT-PCR, as well as other methods, which cannot be performed on encapsulated islets. (C) 2003 Wiley Periodicals, Inc.

Original languageEnglish
Pages (from-to)679-683
Number of pages5
JournalJournal of Biomedical Materials Research. Part A
Volume67A
Issue number2
Publication statusPublished - 1-Nov-2003

    Keywords

  • alginate-poly-L-lysine-alginate microcapsules, islets of Langerhans, trypsin, EDTA, RT-PCR, PROPIDIUM IODIDE, ACRIDINE-ORANGE, BIOCOMPATIBILITY, VIABILITY, GRAFTS, CELLS

ID: 4143238