Characterization of neuronal differentiation in human adipose-derived stromal cells: morphological, molecular, and ultrastructural insights

J Neurosci Methods. 2024 Dec:412:110296. doi: 10.1016/j.jneumeth.2024.110296. Epub 2024 Sep 30.

Abstract

Objective: Adipose-derived stromal cells (ADSCs) have shown promise as a potential source of neural differentiation. In this study, we investigated the morphological, molecular and ultrastructural features of ADSCs during neuronal differentiation.

Methods: ADSCs were induced in vitro and their differentiation was examined at different time points. Immunocytochemical staining was performed to detect the expression of neuron-specific markers NSE and MAP-2. Immunofluorescence double labeling and Western blot detected the co-expression of presynaptic markers (CaMKII, SynCAM1, SYN) and postsynaptic markers (PSD-95, Synapsin I). Scanning electron microscopy (SEM) was performed to detect the synaptic structural features of differentiated neurons.

Results: ADSCs showed diverse morphological features during differentiation, gradually acquiring a neuron-like spindle shape and organized arrangement. The expression of neuron-specific markers and synaptic markers peaked at 5 h of induction. Scanning electron microscopy showed polygonal protrusions of ADSC-derived neurons, and transmission electron microscopy showed characteristic ultrastructures such as nidus, synaptic vesicle-like structures, and tight junctions.

Conclusion: Our findings suggest that ADSCs differentiated for 5 h have neuronal features, including morphological, molecular, and ultrastructural resemblance to neurons, as well as the formation of synaptic structures. These insights contribute to a better understanding of ADSC-based neuronal differentiation and pave the way for future applications in regenerative medicine and neurodegenerative diseases.

Keywords: adipose-derived stromal cells; induction; neurons; release function; synapses; ultrastructure.

MeSH terms

  • Adipose Tissue* / cytology
  • Cell Differentiation* / physiology
  • Cells, Cultured
  • Humans
  • Microscopy, Electron, Scanning
  • Microscopy, Electron, Transmission
  • Neurons* / cytology
  • Neurons* / metabolism
  • Neurons* / ultrastructure
  • Stromal Cells* / metabolism
  • Stromal Cells* / ultrastructure