Direct conversion of adult fibroblasts into motor neurons

Highlights

• Detailed protocol to generate MNs from human adult fibroblasts

• Protocol assesses the identity and efficiency of MN generation in vitro

• Descriptions of troubleshooting common problems during the reprogramming process

Summary

Generation of patient-derived neurons provides an unprecedented approach in modeling neurological diseases. Here, we describe the direct conversion of adult fibroblasts into motor neurons via lentiviral delivery of transcription factors. Compared with iPSC-based approach, directly converted neurons from donors retain features associated with age, making them ideal systems for modeling age-related neurological diseases. Low yield is the major challenge of this protocol. High quality lentiviruses and optimized cell culture conditions are critical to improve the final yield.

For complete details on the use and execution of this protocol, please refer to Ding et al. (2020), Ding et al. (2021), and Liu et al. (2016)