Establishment of TH-EGFP human embryonic stem cell line for specific labeling of dopaminergic neurons

Stem Cell Res. 2024 Dec:81:103613. doi: 10.1016/j.scr.2024.103613. Epub 2024 Nov 17.

Abstract

Dopaminergic (DA) neurons play critical roles in various neurological processes and disorders, particularly Parkinson's disease. To enable precise visualization and tracking of DA neurons, we generated TH-EGFP, a tyrosine hydroxylase (TH)-driven enhanced green fluorescent protein (EGFP)-expressing knock-in cell line, by employing CRISPR/Cas9 technology. We introduced EGFP into the targeted genomic region of human embryonic stem cells (hESCs) and successfully established a TH-EGFP hESC line. Differentiation of TH-EGFP hESCs into human midbrain organoids confirmed the accurate integration of EGFP into TH-positive cells. The TH-EGFP hESC line serves as a valuable reporter for studying the development, maturation, and function of DA neurons.

MeSH terms

  • CRISPR-Cas Systems
  • Cell Differentiation
  • Cell Line
  • Dopaminergic Neurons* / cytology
  • Dopaminergic Neurons* / metabolism
  • Green Fluorescent Proteins* / genetics
  • Green Fluorescent Proteins* / metabolism
  • Human Embryonic Stem Cells* / cytology
  • Human Embryonic Stem Cells* / metabolism
  • Humans
  • Tyrosine 3-Monooxygenase / genetics
  • Tyrosine 3-Monooxygenase / metabolism

Substances

  • Green Fluorescent Proteins
  • enhanced green fluorescent protein
  • Tyrosine 3-Monooxygenase