Proteomic Characterization of Human Neural Stem Cells and Their Secretome During in vitro Differentiation

Frontiers in Cellular Neuroscience, 2021 · DOI: 10.3389/fncel.2020.612560 · Published: January 28, 2021

Regenerative Medicine Neurology Bioinformatics

Simple Explanation

Cell therapies are a promising approach for neurodegenerative diseases and spinal cord injuries. Grafted cells can integrate into damaged tissue, modulate inflammation, or support neuron survival. Characterizing cells before transplantation is crucial to prevent risks like tumorous growth. This study characterized changes in the proteome and secretome of human neural stem cells (NSCs) during differentiation. The study differentiated NSCs spontaneously and with trophic support using BDNF/GDNF supplementation. The researchers used LC-MS/MS to quantify almost three thousand cellular proteins and Luminex multiplex immunoassay to analyze the secretome. This revealed substantial protein differences in the early stages of NSC differentiation.

Study Duration

28 days

Participants

Human Neural Stem Cells (NSCs) derived from the H9 ESC line

Evidence Level

Not specified

Key Findings

1
Substantial protein differences were identified in the early stages of NSC differentiation. More than a third of all the proteins were regulated, including known neuronal and NSC multipotency markers.
2
BDNF/GDNF support affected more the later stages of the NSC differentiation. Pathways activated during differentiation included the HIF-1, Wnt, and VEGF signaling pathways.
3
Secretome analysis revealed significant changes in the secretion of VEGF and IL-6 during NSC differentiation. Increased expression of neuropilin-1 and catenin β-1, participating in VEGF signaling regulation, was observed. VEGF-A isoform 121 induced proliferation and supported survival.

Research Summary

This study provides a comprehensive proteomic and secretomic characterization of human neural stem cells (NSCs) during in vitro differentiation. NSCs were differentiated spontaneously or with trophic support by BDNF/GDNF supplementation, and changes in their proteome and secretome were analyzed. The results revealed substantial protein differences in the early stages of NSC differentiation, with more than a third of all the proteins regulated. BDNF/GDNF support affected more the later stages of differentiation, and key pathways like HIF-1, Wnt, and VEGF signaling were activated. Secretome analysis showed significant changes in VEGF and IL-6 secretion during NSC differentiation, with VEGF-A isoform 121 inducing proliferation and supporting survival. These findings contribute to understanding the regulatory networks behind NSC behavior in nervous system development, tissue regeneration, and cancer.

Practical Implications

Cell-Based Therapies

The study provides insights into the proteomic and secretomic profiles of NSCs, which can be used to optimize cell-based therapies for neurodegenerative diseases and spinal cord injuries.

Targeted Therapies

Identifying key pathways like VEGF, Wnt, and HIF-1 signaling provides potential targets for developing therapies that promote neuronal survival and regeneration.

Understanding Stem Cell Behavior

Elucidating the molecular mechanisms behind NSC proliferation, survival, and differentiation can help in understanding stem cell behavior in nervous system development, tissue regeneration, and cancer.

Study Limitations

1
The H9 NSC line used in the study may not fully represent true multipotent NSCs due to preferential differentiation into neurons.
2
Proteomic and transcriptomic data showed discrepancies, possibly due to differential regulation at mRNA and protein levels.
3
The study was conducted in vitro, and the results may not fully translate to in vivo conditions.

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