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Microbiology JoVE (Open Access) Citable · DOI

Ex Vivo Infection of Live Tissue with Oncolytic Viruses

DOI: 10.3791/2854-v
What you'll learn
  • ✓ Process fresh patient tissue for ex vivo infection
  • ✓ Assess tissue viability after processing
  • ✓ Detect GFP and luciferase viral transgene expression
  • ✓ Quantify viral titers by plaque assay
Protocol

Biopharma Insights Oncolytic viruses are promising for cancer therapeutics. The ability to ascertain the infectability of live tissue specimens obtained from patients prior to treatment is a unique advantage of this therapeutic approach. This protocol describes how to process tissues for ex vivo infection with oncolytic virus and subsequent viral quantification.

Difficulty
intermediate
Total time
~4-6 hrs per sample
Model organism
Human patient tissue (ex vivo)
Biosafety
BSL-2

Steps

1
Process tissue for ex vivo infection

Prepare fresh patient tissue samples by sectioning and placing them in appropriate culture conditions for viral infection.

▶ 02:00
2
Assess tissue viability

Determine the viability of processed tissue samples using standard viability assays to ensure suitability for infection.

▶ 03:00
3
Visualize GFP transgene expression

Use fluorescence microscopy to detect GFP expression from the oncolytic virus, indicating successful infection.

▶ 04:48
4
Visualize luciferase expression with IVIS

Employ an in vivo imaging system (IVIS) to detect luciferase expression, confirming viral replication in the tissue.

▶ 05:50
5
Determine viral titers by plaque assay

Perform a plaque assay on infected tissue homogenates to quantify the amount of infectious virus produced.

▶ 06:33
6
Evaluate viability and infectability correlation

Analyze the relationship between tissue viability and viral infectability, interpreting results for patient-specific therapeutic insights.

▶ 09:52
7
Conclude and interpret protocol results

Summarize the findings and discuss implications for using oncolytic viruses in personalized cancer therapy.

▶ 11:07
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