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. 2023 Mar 6;12(3):bio059845. doi: 10.1242/bio.059845

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© 2023. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 7. — IPS derived NPCs reveal senescence and disrupted stem cell capacity. To confirm our findings about the neural precursor pool in c-organoids, patient derived iPSCs were directly differentiated into neural stem cells. Obtained patient NSCs were studied by immunofluorescence during expansion and after differentiation by mitogen withdrawal. (A) Schematic representation of the protocol used to differentiate iPSCs into NPC. (B) Representative pictures of iPSCs in expansion (left), NPCs in expansion (middle), and differentiated NPCs after 10 days of mitogen withdrawal (right). (C) Representative pictures of immunofluorescence in NPCs during expansion in vitro. Staining for proliferation marker Ki67, stem cell marker PAX6 and senescence marker P16. A decrease of Ki67 and PAX6 expression was observed in MS organoids compared to control. A strong expression of P16 was detected in PPMS NPCs only. (D) Representative pictures of immunofluorescence in NPCs after differentiation by mitogen withdrawal. Staining for neuronal marker DCX, astroglial marker GFAP and the oligodendroglial lineage marker Olig2 were performed. A decrease of DCX and Olig2, associated with an increase of GFAP expression was detected in MS samples, particularly PPMS.