Direct conversion of adult fibroblasts into motor neurons

STAR Protoc. 2021 Oct 23;2(4):100917. doi: 10.1016/j.xpro.2021.100917. eCollection 2021 Dec 17.

Abstract

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).

Keywords: Cell Biology; Cell culture; Developmental biology; Neuroscience; Tissue Engineering.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Differentiation
  • Cells, Cultured
  • Fibroblasts* / cytology
  • Fibroblasts* / metabolism
  • Genetic Vectors / genetics
  • HEK293 Cells
  • Humans
  • Lentivirus / genetics
  • Motor Neurons* / cytology
  • Motor Neurons* / metabolism
  • Transcription Factors* / genetics
  • Transcription Factors* / metabolism

Substances

  • Transcription Factors