Neural stem cell (NSC) transplantation is a promising therapeutic strategy for neurodegenerative diseases and neurological injury. In addition to testing transplant mediated regeneration and functional recovery in animal models of these neuropathologies, several clinical trials are currently underway to test the therapeutic efficacy of these cells in diseases such as Alzheimer’s and spinal cord injury. Hence, there is an urgent need for non-viral methods to safely engineer clinically viable “neuromimetic cells”–NSCs propagated using three-dimensional in vitro systems that most closely mimic the physiological tissue they are derived from.
Chari et al 2017. Curr. Protoc. Stem Cell Biol. 40:2D.19.1-2D.19.16.
In the February Supplement of Current Protocols in Stem Cell Biology, Dr. Divya Chari and her group describe a series of protocols for:
A) preparation of NSC neurosphere and monolayers using mouse brain B) magnetofection–gene delivery using magnetic nanoparticles (MNPs).
Neurospheres offer key advantages in terms of neural graft survival after transplantation while magnetofection enhances gene delivery making this a safe and effective method to generate NSCs for translational applications. The authors describe gene delivery to NSCs using NeuroMag, commercially available transfection-grade MNPs, but the approach can be easily adapted to other MNP formulations, including those used for magnetolabeling (e.g., for non-invasive cell tracking by MRI). The unit details critical parameters that should be addressed when adapting these protocols and addresses procedural safety evaluation methods—a primary concern when manipulating NSC transplant populations. Expected results and troubleshooting tips make this protocol a valuable resource for laboratories engaged in stem cell transplantation research.
In summary, this protocol is the first to describe the use of oscillating magnetic fields for MNP-mediated gene transfer to NSCs, offering a highly, efficacious, non-viral alternative for genetic modification of the clinically significant neural cell transplant population.
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