Transfection of Rat E14 Spinal Cord NPCs

Rat E14 spinal cord–derived NPCs were transfected using 4D-Nucleofector with the P3 Primary Cell Kit (3 million per cuvette; Lonza). Either 3 μg of Negr1-pcDNA3 + 0.5 μg of Pmax-GFP (Lonza) or 3 μg of empty pcDNA3 backbone (31 (link)) + 0.5 μg of Pmax-GFP were used per electroporation in the 16-well Nucleocuvette Strip and plated on PDL-coated six-well plates, matured in vitro for 6 days, and replated on myelin substrates as described above.

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Poplawski G.H., Lie R., Hunt M., Kumamaru H., Kawaguchi R., Lu P., Schäfer M.K., Woodruff G., Robinson J., Canete P., Dulin J.N., Geoffroy C.G., Menzel L., Zheng B., Coppola G, & Tuszynski M.H. (2018). Adult rat myelin enhances axonal outgrowth from neural stem cells. Science translational medicine, 10(442), eaal2563.

Publication 2018

4D-Nucleofector P3 Primary Cell Kit Pmax-GFP

Backbone Cell Electroporation Myelin Spinal cord

Corresponding Organization : United States Department of Veterans Affairs

Other organizations : University of California, Los Angeles, University Medical Center of the Johannes Gutenberg University Mainz, Johannes Gutenberg University Mainz

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Variable analysis

independent variables

Negr1-pcDNA3 transfection
Empty pcDNA3 backbone transfection

dependent variables

Maturation of rat E14 spinal cord-derived NPCs in vitro for 6 days
Replating of NPCs on myelin substrates

control variables

Cell density (3 million per cuvette)
Pmax-GFP co-transfection (0.5 μg)

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