( ...)Effectene Transfection Reagent provides efficient gene delivery ,,, to primary neuronal cell cultures

Jochen Meier, Ivonne Strmel, Radu Iosub, Sonja Schmidt, and Rosemarie Grantyn
Developmental Physiology, Johannes Mller Institute, Humboldt University Medical School (Charit), Berlin, Germany

Efficient delivery of DNA to primary neuronal cell cultures is of critical importance for extending our knowledge of the molecular basis of neuronal physiology. Here, we describe the use of Effectene Transfection Reagent for successful transfection of primary differentiated neurons in rat superior colliculus cultures.

Enhanced green fluorescent protein (EGFP) chimeras have proven to be a powerful tool for studying the underlying cellular mechanisms for dynamic remodeling of synaptic compounds in living neurons (1). However, a major obstacle to the use of such chimeras is an effective gene delivery method. Gene delivery based on non-viral DNA vehicles is an attractive alternative because it can be applied under a wide range of laboratory conditions. We developed a transfection procedure using Effectene Reagent and found it highly suited for easy and efficient transfection of differentiated neurons in primary cultures from rat superior colliculus. Effectene Reagent shows low cytotoxicity and delivers efficient transfection using small amounts of DNA. Furthermore, transfection can be carried out in the presence of serum.

Materials and methods

Superior colliculus neurons were prepared from embryonic day 19 Wistar rats, and cells were plated at a density of 7.5 x 104 cells/cm2 on laminin-coated glass coverslips (16 mm). Cultures were maintained in neurobasal B27 medium supplemented with 1% fetal calf serum. After neuron attachment, coversli
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