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Updated: Dec 27, 2025
Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles
Published on: April 6, 2013
Olig2 positive cells derived from hair follicle neural crest stem cells in rats
Tao Wang1, Haiyan Lin1, Fang Liu1
1Department of Anatomy, the Second Military Medical University/Naval Medical University, China.
Abstract:
Motor neuron disease (MND) is a kind of common clinical nervous system disease with typical characteristic of progressive motor neurons degeneration or death. Motor neuron derived from stem cells or motor neuron progenitor cells will be a good choice to be used for treatment of the disease. In this study, we used the combination of 5 small molecular including CHIR99021 (CHIR), SB431542 (SB), DMH1 (DMH), retinoic acid (RA) and Purmorphamine (Pur) to induce hair follicles neural crest stem cells (hfNCSCs) to motor neurons progenitors (MNPs). Valproic acid (VPA) was used to make MNPs proliferation. RA and Pur were used to try to induce MNPs toward motor neurons (MNs) and CpdE was tried for MNs maturation. Nestin, β-tubulin Ш (Tuj1), microtubule associated protein 2 (MAP2), Olig2, choline acetyltransferase (ChAT)and TUBB3 were examined at protein and mRNA levels by immunofluoresence cytochemistry, western blot and real time PCR at 6, 16 and 22 days. Our data showed cells changed into bipolar or multipolar shape forming the cell clusters like scattered rosettes. Nestin expression decreased significantly at 22 days. Compared to 6 days, percentage of Olig2 + MNPs was higher, (88.53 ± 6.67)%, and Olig2 expression at protein and gene level was lower at 22 days. Percentage of MAP2 positive cells increased to (90.62 ± 2.31) % and ChAT positive cells increased to (83.29 ± 6.62) % at 22 days. But no expression of ChAT was examined by western blot and real time PCR. It indicates that these 5 molecular can differentiate hfNCSCs into Olig2 positive cells with a unipotent differentiation toward motor neurons.
Insights
Researchers used five small molecules to transform hair follicle stem cells into motor neuron progenitors. These cells showed potential for motor neuron differentiation, offering a promising avenue for treating motor neuron diseases.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Regenerative Medicine
Background:
- Motor neuron disease (MND) involves progressive degeneration of motor neurons.
- Generating motor neurons from stem cells is a potential therapeutic strategy for MND.
- Hair follicle neural crest stem cells (hfNCSCs) offer a source for cell-based therapies.
Purpose of the Study:
- To investigate the differentiation of hfNCSCs into motor neuron progenitors (MNPs) and subsequently motor neurons (MNs) using a specific combination of small molecules.
- To evaluate the expression of key neuronal markers during the differentiation process.
- To assess the potential of these induced cells for treating motor neuron diseases.
Main Methods:
- Induction of hfNCSCs into MNPs using CHIR99021, SB431542, DMH1, retinoic acid (RA), and Purmorphamine (Pur).
- Proliferation of MNPs using Valproic acid (VPA).
- Induction towards MNs and maturation using RA, Pur, and CpdE.
- Analysis of Nestin, Tuj1, MAP2, Olig2, ChAT, and TUBB3 expression at protein and mRNA levels via immunofluorescence, Western blot, and real-time PCR.
Main Results:
- Observed morphological changes in cells, including bipolar/multipolar shapes and rosette-like clusters.
- Significant decrease in Nestin expression by day 22.
- Increased percentage of Olig2+ MNPs, but decreased Olig2 protein and gene expression at day 22.
- Increased MAP2 and ChAT positive cell percentages by day 22.
- ChAT expression was not detected via Western blot or real-time PCR, suggesting unipotent differentiation.
Conclusions:
- The combination of five small molecules can effectively induce hfNCSCs into Olig2+ cells.
- These cells demonstrate a unipotent differentiation potential towards motor neurons.
- This method provides a foundation for developing cell-based therapies for motor neuron diseases.
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