Comparative transfection studies of human ovarian carcinoma cells in vitro, ex vivo and in vivo with poly(2-(dimethylamino)ethyl methacrylate)-based polyplexes
✍ Scribed by Petra van de Wetering; Nancy M.E. Schuurmans-Nieuwenbroek; Wim E. Hennink; Gert Storm
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 169 KB
- Volume
- 1
- Category
- Article
- ISSN
- 1099-498X
No coin nor oath required. For personal study only.
✦ Synopsis
Background Poly(2-(dimethylamino)ethyl methacrylate) (p(DMAEMA)) can be used successfully for in vitro transfection of different cell lines, including the OVCAR-3 human ovarian carcinoma cell line. The aim of this study was to investigate whether it is possible to transfect OVCAR-3 cells in vivo with polyplexes containing p(DMAEMA).
Methods In order to understand the generally observed gap between in vitro and in vivo transfection, we gradually went from in vitro to in vivo transfection of OVCAR-3 cells, while keeping the exposure conditions the same, as far as possible. To ®nd the reason for the negligible degree of in vivo transfection, in vitro cultured OVCAR-3 cells were transfected in the presence of peritoneal ascites ¯uid. Next, the in¯uence of hyaluronic acid, one of the ascites components, on the transfection ef®ciency was studied.
Results P(DMAEMA)-containing polyplexes can transfect OVCAR-3 cells in vitro with an overall transfection ef®ciency of 10%. Cells grown in vivo can be transfected ex vivo with p(DMAEMA)/plasmid complexes with an overall transfection ef®ciency of ,1±2%. When transfection complexes are injected i.p. into nude mice bearing OVCAR-3 cells in the peritoneal cavity, the degree of in vivo transfection ef®ciency achieved is negligible.
In vitro cultured OVCAR-3 cells were also transfected with polyplexes in the presence of peritoneal ascites ¯uid. The results indicate that one or more components of ascites had a negative effect on the transfection ef®ciency of p(DMAEMA)-containing polyplexes. To elucidate which component(s) of ascites may have interfered, the in¯uence of hyaluronic acid, one of the ascites components, on the transfection ef®ciency was studied. The outcome suggests that hyaluronic acid may have induced a negative effect on the transfection capability of p(DMAEMA)-containing polyplexes.
Conclusion P(DMAEMA) is an ef®cient transfectant in vitro and ex vivo. However, transfected cells were not detected in vivo which may be caused by a negative in¯uence of components of the ascites ¯uid.