Journal
MOLECULAR THERAPY
Volume 3, Issue 1, Pages 113-121Publisher
ACADEMIC PRESS INC
DOI: 10.1006/mthe.2000.0235
Keywords
organotypic; electroporation; lipotransfection; biolistics; gene transfer; cerebellum
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Funding
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [R42NS038002, R41NS038002] Funding Source: NIH RePORTER
- NINDS NIH HHS [NS38002] Funding Source: Medline
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Organotypic slice cultures from postnatal day 12 mouse cerebellum were transfected using three nonviral methods: biolistics (gene gun), lipotransfection, and electroporation. The plasmid transferred, pHD17-25Q-GFP, encoded a fusion protein with a green fluorescent protein (GFP) component. Optimal conditions for both lipotransfection and electroporation are the same as those previously found in live animal models. Electroporation (26 +/- 6) and biolistics (34 +/- 4.4) provide a better rate of transfer than lipotransfection (15 +/- 2.2) in slice cultures and are comparable to each other. Each of the transfer methods produced positive signals in a heterogeneous population of glial and neuronal cells. These data provide a base for optimal transfection of slice cultures, allowing the development of therapeutic constructs, and support the idea that successful refinement of nonviral delivery methods for in vivo use is possible using brain slice cultures.
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