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Single-Step Plasmid Based Reprogramming of Human Dermal Fibroblasts to Induced Neural Stem Cells

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Single-Step Plasmid Based Reprogramming of Human Dermal Fibroblasts to Induced Neural Stem Cells

Luis Azmitia et al. Methods Mol Biol.

Abstract

Reprogramming of somatic cells to induced pluripotent stem cells (iPSC) and subsequent differentiation opened up the opportunity of deriving cell types in vitro which (like neurons) had a very restricted accessibility in the past. However, cell culture protocols for iPSC reprogramming, neural induction and differentiation tend to be labor and time intensive, costly and commonly depend on viral vector delivery. Single-step reprogramming to induced neural stem cells (iNSC) avoids many of the necessary intermediate steps of the aforementioned method but yields a cell type that proliferates over longer time spans and readily differentiates to mature neurons when required. Here we describe a plasmid based reprogramming protocol employing defined, commercially available components for induction and proliferation of iNSC, followed by a defined, small molecule based differentiation step toward mature neurons. The described method might be of particular interest for groups with limited resources and/or restricted access to higher biosafety level facilities required for viral transduction, but also for groups requiring a high throughput for dealing with large numbers of cell lines.

Keywords: Direct neural reprogramming; Human dermal fibroblasts; Human neurons; Induced neural stem cells; Plasmid transfection; Small molecule based differentiation; Small molecules.

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