The organotypic multicellular spheroid is a relevant three-dimensional model to study adenovirus replication and penetration in human tumors in vitro

Jacques Grill, Martine L.M. Lamfers, Victor W. Van Beusechem, Clemens M. Dirven, D. Shareen Pherai, Mathijs Kater, Paul Van Der Valk, Ronald Vogels, W. Peter Vandertop, Herbert M. Pinedo, David T. Curiel, Winald R. Gerritsen

Research output: Contribution to journalArticlepeer-review

63 Scopus citations

Abstract

The use of adenoviruses for gene transfer and as oncolytic agents is currently receiving wide-spread attention. As specific constraints to adenovirus distribution and spread cannot be studied in cell cultures, there is a need for an in vitro three-dimensional (3D) model mimicking the in vivo biology of tumors. We studied the interactions between tumor and adenoviruses using multicellular spheroids grown from primary brain tumor material. Using β-galactosidase and luciferase reporter genes expressed by replication-defective adenoviruses, we showed that infection was restricted to the first layer of cells. Using a replication-competent adenovirus expressing the luciferase gene, was showed that transgene expression in the spheroid was considerably enhanced and that viral spreading deep into the 3D structure took place. In addition, a tetrazolium salt-based metabolic assay could be used to compare the oncolytic activity of different concentrations of replication-competent adenoviruses. We can conclude that organotypic spheroids offer a versatile in vitro system for studying distribution, spread, and oncolysis by adenoviruses in a clinically relevant model.

Original languageEnglish
Pages (from-to)609-614
Number of pages6
JournalMolecular Therapy
Volume6
Issue number5
DOIs
StatePublished - Nov 1 2002

Keywords

  • Adenovirus
  • Cancer gene therapy
  • In vitro model
  • Oncolytic viruses
  • Replication
  • Spheroid

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