Geer MAV, Kuhlmann KF, Bakker CT, Kate FJT, Elferink RPO, Bosma PJ. Ex-vivo evaluation of gene therapy vectors in human pancreatic (cancer) tissue slices. World J Gastroenterol 2009; 15(11): 1359-1366 [PMID: 19294766 DOI: 10.3748/wjg.15.1359]
Corresponding Author of This Article
Dr. Piter J Bosma, Liver Center, Academic Medical Center of the University of Amsterdam, Meibergdreef 69/71, 1105 BK Amsterdam, The Netherlands. p.j.bosma@amc.uva.nl
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Brief Articles
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World J Gastroenterol. Mar 21, 2009; 15(11): 1359-1366 Published online Mar 21, 2009. doi: 10.3748/wjg.15.1359
Ex-vivo evaluation of gene therapy vectors in human pancreatic (cancer) tissue slices
Michael A van Geer, Koert FD Kuhlmann, Conny T Bakker, Fibo JW ten Kate, Ronald PJ Oude Elferink, Piter J Bosma
Michael A van Geer, Conny T Bakker, Ronald PJ Oude Elferink, Piter J Bosma, Liver Center, Academic Medical Center of the University of Amsterdam, Meibergdreef 69/71, 1105 BK Amsterdam, The Netherlands
Koert FD Kuhlmann, Department of Surgery, Academic Medical Center of the University of Amsterdam, Meibergdreef 9, 1105BK Amsterdam, The Netherlands
Fibo JW ten Kate, Department of Pathology, Academic Medical Center of the University of Amsterdam, Meibergdreef 9, 1105 BK Amsterdam, The Netherlands
Author contributions: van Geer MA generated the Adenovirus preparations and performed the transduction experiments in the slices; Kuhlmann KFD provided the resection specimens and performed the immune histological stainings; Bakker CT produced the Lentivirus and part of the adenovirus preparations and performed the slicing of the resection specimens; ten Kate FJW scored the viability of the slices after ex-vivo culture; Oude Elferink RPJ designed the study and obtained the grant required for performing this study and corrected the paper; Bosma PJ designed the study, generated the AAV vector and wrote the paper.
Correspondence to: Dr. Piter J Bosma, Liver Center, Academic Medical Center of the University of Amsterdam, Meibergdreef 69/71, 1105 BK Amsterdam, The Netherlands. p.j.bosma@amc.uva.nl
Telephone: +31-20-5668850
Fax: +31-20-5669190
Received: December 26, 2008 Revised: February 23, 2009 Accepted: March 2, 2009 Published online: March 21, 2009
Abstract
AIM: To culture human pancreatic tissue obtained from small resection specimens as a pre-clinical model for examining virus-host interactions.
METHODS: Human pancreatic tissue samples (malignant and normal) were obtained from surgical specimens and processed immediately to tissue slices. Tissue slices were cultured ex vivo for 1-6 d in an incubator using 95% O2. Slices were subsequently analyzed for viability and morphology. In addition the slices were incubated with different viral vectors expressing the reporter genes GFP or DsRed. Expression of these reporter genes was measured at 72 h after infection.
RESULTS: With the Krumdieck tissue slicer, uniform slices could be generated from pancreatic tissue but only upon embedding the tissue in 3% low melting agarose. Immunohistological examination showed the presence of all pancreatic cell types. Pancreatic normal and cancer tissue slices could be cultured for up to 6 d, while retaining viability and a moderate to good morphology. Reporter gene expression indicated that the slices could be infected and transduced efficiently by adenoviral vectors and by adeno associated viral vectors, whereas transduction with lentiviral vectors was limited. For the adenoviral vector, the transduction seemed limited to the peripheral layers of the explants.
CONCLUSION: The presented system allows reproducible processing of minimal amounts of pancreatic tissue into slices uniform in size, suitable for pre-clinical evaluation of gene therapy vectors.
