Entrapment of retroviral vector producer cells in three-dimensional alginate scaffolds for potential use in cancer gene therapy
✍ Scribed by Mona Dvir-Ginzberg; Alexander Konson; Smadar Cohen; Riad Agbaria
- Publisher
- John Wiley and Sons
- Year
- 2006
- Tongue
- English
- Weight
- 252 KB
- Volume
- 80B
- Category
- Article
- ISSN
- 1552-4973
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✦ Synopsis
Abstract
Abstract: We explored the possibility of entrapping retroviral vector producing cells (VPC) within porous 3D matrix to induce a local and sustained release of viral particles to the malignant milieu. PA317/STK, which constantly shed reroviral vectors, was used to transduce cancer cells with the herpes simplex virus thymidine kinase (HSV‐tk) gene. Once HSV‐tk is expressed, it preferentially phosphorylates nucleoside analog prodrugs, such as ganciclovir (GCV) and N‐methanocarbathymidine (N‐MCT), to their active triphosphate metabolites, which when incorporated into cellular DNA cause cell death. PA317/STK cells were seeded within 3D alginate scaffold at two different cell densities via static seeding procedure. In vitro assays determined that PA317/STK seeded at high‐cell density in scaffolds maintained constant cell number, low cell leakage, and spheroid morphology with viral vector transfection activity. Postcell‐seeding viral vector activity was confirmed by transfection of murine colon cancer cells (MC38) with conditioned media originated from VPC‐containing scaffolds and the subsequent ability to generate N‐MCT triphosphate. Preliminary in vivo transplantation of VPC‐containing scaffolds into the peritoneal cavity of mice bearing intraperitoneal MC38 tumors with 2 weeks subsequent GCV administration resulted in a significantly higher survival rate relative to control groups. Our results demonstrate the feasibility of employing alginate scaffolds to efficiently entrap and support PA317/STK cells for cancer gene therapy. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2007