Successful incorporation of short-interfering RNA into islet cells by in situ perfusion

S. P. Bradley, T. F. Kowalik, Cristiana Rastellini, M. A. Da Costa, A. B. Bloomenthal, Luca Cicalese, G. P. Basadonna, M. E. Uknis

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Islet transplantation offers a potential cure for type I diabetes, although its success has been limited, due to loss of cells by apoptosis stimulated by the procurement, ischemia, and the isolation process itself. RNA interference (RNAi) as mediated by short interfering RNAs (siRNAs) has become a potent tool to manipulate gene expression in mammalian cells. We describe the first successful introduction of siRNA directly into pancreatic islet cells both during in situ perfusion and from intravenous tail vein injection (in vivo). Methods. siRNA was targeted to the pancreatic islets of BALB/c mice by retrograde portal vein perfusion or tail vein injection. Cy3-labeled siRNA was dissolved in University of Wisconsin (UW) solution at 2 μg/mL. After delivery pancreata were placed in cold storage at 4°C in UW solution for 24 hours, followed by processing for immunofluorescent staining for insulin. Fluorescent imaging was obtained using a Nikon DIAPHOT 300 Inverted Micoscope with a Zeiss AxioCam and OpenLab image capturing software. Results. In situ delivery of siRNA was demonstrated by fluorescent imaging composites of (red) siRNA in and along (green) insulin stained islets from pancreas sections as compared with untreated control sections. The siRNA was detected mainly in and along venous structures throughout the pancreatic tissue. In vivo delivery of siRNA into islets was observed by fluorescent images taken of isolated islets in culture. Conclusions. We have described the successful delivery of siRNA to pancreatic islets via a novel in situ pancreas perfusion technique and in vivo delivery via tail vein injection.

Original languageEnglish (US)
Pages (from-to)233-236
Number of pages4
JournalTransplantation Proceedings
Volume37
Issue number1
DOIs
StatePublished - Jan 2005
Externally publishedYes

Fingerprint

Islets of Langerhans
Small Interfering RNA
Perfusion
Tail
Pancreas
Veins
Injections
Insulin
Islets of Langerhans Transplantation
Portal Vein
RNA Interference
Type 1 Diabetes Mellitus
Software
Ischemia
Apoptosis
Staining and Labeling
Gene Expression

ASJC Scopus subject areas

  • Surgery
  • Transplantation

Cite this

Bradley, S. P., Kowalik, T. F., Rastellini, C., Da Costa, M. A., Bloomenthal, A. B., Cicalese, L., ... Uknis, M. E. (2005). Successful incorporation of short-interfering RNA into islet cells by in situ perfusion. Transplantation Proceedings, 37(1), 233-236. https://doi.org/10.1016/j.transproceed.2004.12.181

Successful incorporation of short-interfering RNA into islet cells by in situ perfusion. / Bradley, S. P.; Kowalik, T. F.; Rastellini, Cristiana; Da Costa, M. A.; Bloomenthal, A. B.; Cicalese, Luca; Basadonna, G. P.; Uknis, M. E.

In: Transplantation Proceedings, Vol. 37, No. 1, 01.2005, p. 233-236.

Research output: Contribution to journalArticle

Bradley, SP, Kowalik, TF, Rastellini, C, Da Costa, MA, Bloomenthal, AB, Cicalese, L, Basadonna, GP & Uknis, ME 2005, 'Successful incorporation of short-interfering RNA into islet cells by in situ perfusion', Transplantation Proceedings, vol. 37, no. 1, pp. 233-236. https://doi.org/10.1016/j.transproceed.2004.12.181
Bradley, S. P. ; Kowalik, T. F. ; Rastellini, Cristiana ; Da Costa, M. A. ; Bloomenthal, A. B. ; Cicalese, Luca ; Basadonna, G. P. ; Uknis, M. E. / Successful incorporation of short-interfering RNA into islet cells by in situ perfusion. In: Transplantation Proceedings. 2005 ; Vol. 37, No. 1. pp. 233-236.
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