Abstract

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Background and Purpose

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This study aimed to evaluate glial cell differentiation within a 3D central nervous system (CNS) model using induced pluripotent stem cell (iPSC)-derived neurons. The goal was to determine optimal growth factor formulations to promote astrocyte, oligodendrocyte precursor cell (OPC), and oligodendrocyte differentiation.

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Methods

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The research team tested five different media formulations over a 12-week period. They assessed differentiation markers using immunohistochemistry (IHC), quantitative PCR (qPCR), and Western blot techniques. Markers such as GFAP (astrocytes), PDGFRα (OPCs), and Olig2 (oligodendrocytes) were tracked across multiple time points.

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Results

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• Astrocyte Differentiation: Test 5 showed the highest GFAP expression, indicating robust astrocyte growth. Expression steadily increased between day 56 and day 84.
• OPC and Oligodendrocyte Differentiation: Test 5 exhibited a peak in OPC and non-myelinating oligodendrocyte markers at day 70. Key markers like PDGFRα and NG2 were notably elevated.
• Spheroid Growth: Spheroids in Test 5 showed increased diameter over time with consistent circularity.

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Conclusion

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Test 5 proved the most effective formulation, fostering enhanced glial differentiation across astrocytes, OPCs, and oligodendrocytes. Future studies will focus on investigating demyelinating and promyelinating drug effects on oligodendrocyte development.