Protocol for 3D direct reprogramming of human glia into dopaminergic neurons with gene expression and immunocytochemistry validation
Year: 2025
Type: article
Abstract: Direct reprogramming of somatic cells into neurons offers a promising strategy for studying neurological disorders and developing cell-based therapies. This protocol describes the 3D direct reprogramming of human glial progenitor cells into dopaminergic neurons. It includes spheroid formation, lentiviral transduction, neuronal induction using doxycycline, and validation via gene expression and immunocytochemistry. This versatile approach enables efficient reprogramming of human cells into dopaminergic neurons. For complete details on the use and execution of this protocol, please refer to Giacomoni et al. 1 • Steps for thawing and expanding human glial progenitor cells before reprogramming • Instructions for co-seeding glial cells with lentivirus to initiate 3D spheroid formation • Guidance on applying media changes to transduced spheroids to induce neuronal conversion • Procedures for validating neuronal identity using gene expression and immunostaining Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Direct reprogramming of somatic cells into neurons offers a promising strategy for studying neurological disorders and developing cell-based therapies. This protocol describes the 3D direct reprogramming of human glial progenitor cells into dopaminergic neurons. It includes spheroid formation, lentiviral transduction, neuronal induction using doxycycline, and validation via gene expression and immunocytochemistry. This versatile approach enables efficient reprogramming of human cells into dopaminergic neurons.
Source: STAR Protocols
Institution: Lund University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W4414168703