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image of Innovative Approaches to Neural Differentiation: Chondroitin 4-Sulfate and Chondroitinase Induce Differentiation in Human-induced Pluripotent Stem Cells

Abstract

Background

Stem cells have recently gained prominence in regenerative medicine, particularly in the treatment of neurological disorders. As a result, Human-induced Pluripotent Stem Cells (hiPSCs) have become a significant focus.

Objective

This study aimed to differentiate hiPSCs into neural lineages under conditions using forskolin and retinoic acid in an induction medium combined with chondroitin 4-sulfate and chondroitinase.

Methods

Optimal component concentrations were determined using the MTT assay and acridine orange/ethidium bromide (AO/EB) staining. Subsequently, neural-specific genes (, , , , and ) and proteins (gamma enolase, MAP-2, and β-tubulin III) were assessed using Real-time PCR analysis and immunofluorescence staining to provide a comprehensive evaluation of differentiated cells.

Results

Our study demonstrated a significant enhancement in neural-specific gene and protein markers during the 7th and 14th days of differentiation in the presence of combined chondroitin 4-sulfate and chondroitinase, demonstrating a higher efficacy compared with the application of isolated enzymes or substrates.

Conclusion

These findings emphasize the potential importance of chondroitin 4-sulfate and chondroitinase as important factors in promoting the neural differentiation of hiPSCs. It seems that chondroitin 4-sulfate may activate cellular signaling pathways that are effective in inducing neural differentiation. Our findings in this research provide new opportunities to advance regenerative therapies for neurological disorders.

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2025-06-19
2025-09-14

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/content/journals/cscr/10.2174/011574888X365326250610113501
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Innovative Approaches to Neural Differentiation: Chondroitin 4-Sulfate and Chondroitinase Induce Differentiation in Human-induced Pluripotent Stem Cells
Bentham Science Publishers ; https://doi.org/10.2174/011574888X365326250610113501
/content/journals/cscr/10.2174/011574888X365326250610113501
/content/journals/cscr/10.2174/011574888X365326250610113501
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  • Article Type:     Research Article
Keywords: neural differentiation ; chondroitin 4-sulfate ; Chondroitinase ; hiPSCs

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