Topic
Tissue Slice Technology
About: Tissue Slice Technology is a research topic. Over the lifetime, 4 publications have been published within this topic receiving 74 citations.
Papers
TL;DR: A clinically relevant tissue slice technology is presented to preclinically test improved vector characteristics to determine improved characteristics of such agents.
Abstract: Background Inefficiency, aspecificity and toxicity of gene transfer vectors hamper gene therapy from showing its full potential. On this basis significant research currently focuses on developing vectors with improved infection and/or expression profiles. Screening assays with validity to the clinical context to determine improved characteristics of such agents are not readily available since this requires a close relationship to the human situation. We present a clinically relevant tissue slice technology to preclinically test improved vector characteristics. Methods Slices were prepared from rat, mouse and human liver samples and from tumor tissue. Specificity of gene expression and replication was determined by infecting target and non-target tissue slices with transcriptionally retargeted adenoviruses and oncolytic viruses. Results Using rat liver slices, we demonstrate efficient knob-mediated adenoviral infectivity. A favorable tumor-on/liver-off profile, resembling in vitro and mouse in vivo data, was shown for a tumor-specific transcriptionally retargeted adenovirus by infecting slices prepared from tumor or liver tissue. Similar liver-off data were found for mouse, rat and human samples (over 3-log lower activity of the tumor-specific promoter compared to cytomegalovirus (CMV)). More importantly, we show that this technology when applied to human livers is a powerful tool to determine aspecific replication of oncolytic viruses in liver tissue. A 2- to 6-log reduction in viral replication was observed for a tumor-specific oncolytic virus compared to the wild-type adenovirus. Conclusions The precision-cut tissue slice technology is a powerful method to test specificity and efficiency of gene transfer as well as of viral replication using human tissue. Copyright (c) 2005 John Wiley & Sons, Ltd.
40 citations
TL;DR: 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.
Abstract: AIM: To culture human pancreatic tissue obtained from small resection specimens as a pre-clinical model for examining virus-host interactions. 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.
32 citations
TL;DR: The results of this investigation demonstrate the potential toxicologic utility of combining in vitro tissue‐slice technology with high‐resolution 2‐DE protein mapping and offers a novel approach for toxicity screening and testing, reduces experimental cost, and reduces the use of laboratory animals.
Abstract: Advances in tissue slice technology and a recent novel application of this technique to reproductive toxicology using bovine testis have demonstrated the remarkable utility of this approach. The objective of the present study was to combine this in vitro toxicity test system with large-scale two-dimensional polyacrylamide gel electrophoresis (2-DE) to detect and study alterations in testicular-slice protein patterns as molecular correlates of 1,3,5-trinitrobenzene (TNB) and 1,3-dinitrobenzene (DNB) toxicity. Previous studies have shown that testicular slices remain viable for > 24 h and, as measured by protein synthesis inhibition, TNB causes dose-related injury. Tissue-slices were prepared from bovine testicles incubated for 2, 4 or 6 h and exposed to either 100 microM, 500 microM or 1 mM DNB or TNB in the incubation medium. Slices were collected, solubilized, and separated by large scale 2-DE. Resulting protein patterns were then examined by image analysis, which revealed coefficients of variation in protein spot abundance comparable to patterns from fresh rodent tissue samples. Furthermore, specific protein alterations indicated dose-related inductions and declines in protein abundance, some progressive over time. The results of this investigation demonstrate the potential toxicologic utility of combining in vitro tissue-slice technology with high-resolution 2-DE protein mapping. The consolidation of these methods offers a novel approach for toxicity screening and testing, reduces experimental cost, and reduces the use of laboratory animals.
9 citations
1 Jan 2006
TL;DR: A tissue chip based system was created for generating large numbers of samples from a single organ and coupled with fluorescent indicators to maximize detection sensitivity for specific cellular processes and to create an easily deployable method of rapid detection.
Abstract: In vitro toxicology studies are hindered by the use of specific cellular systems which solely examine one cell type. Precision-cut tissue slices mimic specific organ toxicity as normal cellular heterogeneity and organ architecture are retained. Experiments were performed using tissue slices from transgenic mice with enzyme reporter proteins for rapid analysis. CYP 1A1/β-galactosidase transgenic mouse liver and kidney slices challenged with 20 μM BNF for 24 hr remain viable and display organ-specific induction of β-galactosidase (~30-fold in liver and 3-fold in kidney). AP-1/luciferase transgenic mouse tissue slices incubated with 9 μM TPA also remained viable while exhibiting a tissueand time-dependent induction of luciferase. In kidney slices, luciferase induction was approximately 1.5-fold at 2 hr, which increased to 2.5-fold at 4 hr. Liver slices displayed a rapid increase in luciferase at 2 hr (approximately 3-fold) which was abolished at 4 hr. To quicken experimental design via decreased sample preparation, a custom transgenic mouse was created based upon a fluorescent reporter protein. Subsequent studies with slices from this strain and another fluorescent-based transgenic strain did not display reporter induction. For optimization of the smaller tissues of mice and to create an easily deployable method of rapid detection, a tissue chip based system was created for generating large numbers of samples from a single organ and coupled with fluorescent indicators to maximize detection sensitivity for specific cellular processes. Fluorescence of 5-carboxyfluorescein increased at high concentrations of iodoacetamide (IAM), a quick-acting toxicant, indicating disruption of cellular
1 citations
Related Topics (5)
Performance Metrics
| Year | Papers |
|---|---|
| 2009 | 1 |
| 2006 | 2 |
| 1997 | 1 |