Cholinergic and dopaminergic neuronal differentiation of human adipose tissue derived mesenchymal stem cells

J Cell Physiol. 2018 Feb;233(2):936-945. doi: 10.1002/jcp.25937. Epub 2017 Jun 22.

Abstract

Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into various cell types such as cartilage, bone, and fat cells. Recent studies have shown that induction of MSCs in vitro by growth factors including epidermal growth factor (EGF) and fibroblast growth factor (FGF2) causes them to differentiate into neural like cells. These cultures also express ChAT, a cholinergic marker; and TH, a dopaminergic marker for neural cells. To establish a protocol with maximum differentiation potential, we examined MSCs under three experimental culture conditions using neural induction media containing FGF2, EGF, BMP-9, retinoic acid, and heparin. Adipose-derived MSCs were extracted and expanded in vitro for 3 passages after reaching >80% confluency, for a total duration of 9 days. Cells were then characterized by flow cytometry for CD markers as CD44 positive and CD45 negative. MSCs were then treated with neural induction media and were characterized by morphological changes and Q-PCR. Differentiated MSCs expressed markers for immature and mature neurons; β Tubulin III (TUBB3) and MAP2, respectively, showing the neural potential of these cells to differentiate into functional neurons. Improved protocols for MSCs induction will facilitate and ensure the reproducibility and standard production of MSCs for therapeutic applications in neurodegenerative diseases.

Keywords: adipose tissue; cholinergic neurons; dopaminergic neurons; mesenchymal stem cell.

MeSH terms

  • Adipose Tissue / cytology*
  • Adult
  • Cell Lineage
  • Cell Separation
  • Cells, Cultured
  • Cholinergic Neurons / drug effects
  • Cholinergic Neurons / metabolism
  • Cholinergic Neurons / physiology*
  • Culture Media / metabolism
  • Dopaminergic Neurons / drug effects
  • Dopaminergic Neurons / metabolism
  • Dopaminergic Neurons / physiology*
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Humans
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / metabolism
  • Mesenchymal Stem Cells / physiology*
  • Microtubule-Associated Proteins / metabolism
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / metabolism
  • Neural Stem Cells / physiology*
  • Neurogenesis* / drug effects
  • Phenotype
  • Tubulin / metabolism

Substances

  • Culture Media
  • MAP2 protein, human
  • Microtubule-Associated Proteins
  • TUBB3 protein, human
  • Tubulin