A large-scale invitro 3D tumor model was generated to evaluate gene delivery procedures invivo. This 3D tumor model consists of a " tissue-like" spheroid that provides a micro-environment supportive of melanoma proliferation, allowing cells to behave similarly to cells invivo. This functional spheroid measures approximately 1cm in diameter and can be used to effectively evaluate plasmid transfection when testing various electroporation (EP) electrode applicators. In this study, we identified EP conditions that efficiently transfect green fluorescent protein (GFP) and interleukin 15 (IL-15) plasmids into tumor cells residing in the 3D construct. We found that plasmids delivered using a 6-plate electrode applying 6 pulses with nominal electric field strength of 500V/cm and pulse-length of 20ms produced significant increase of GFP (7.3-fold) and IL-15 (3.0-fold) expression compared to controls. This invitro 3D model demonstrates the predictability of cellular response toward delivery techniques, limits the numbers of animals employed for transfection studies, and may facilitate future developments of clinical trials for cancer therapies invivo. © 2012 Elsevier Ltd.
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