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Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for <t>GAP43.</t> Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.
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Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for <t>GAP43.</t> Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.
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Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for <t>GAP43.</t> Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.
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Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for <t>GAP43.</t> Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.
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Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for <t>GAP43.</t> Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.
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Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for GAP43. Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.

Journal: Frontiers in Neuroscience

Article Title: A pH-responsive and self-healing CEC/PEG-(CHO) 2 hydrogel for controlled neuropeptide S delivery and endogenous neural repair after spinal cord injury

doi: 10.3389/fnins.2026.1933351

Figure Lengend Snippet: Phenotypic characterization of mouse NSPCs and the Pro-neurogenic effects of NPS loaded CEC/PEG-(CHO) 2 hydrogel scaffold In Vitro. (a) After 7 days in proliferation medium, isolated NSPCs formed neurospheres. Scale bar = 100 μm. (b–d) Neurospheres were positive for the NSPC markers Nestin, SOX2, and PAX6. Scale bar = 100 μm. (e–g) Quantification of Nestin + , SOX2 + , and PAX6 + cells. (h) Schematic diagram of NSPC isolation. (i) Schematic of the NSPC differentiation assay. (j) After 7 days of differentiation, immature neurons were immunostained for Tuj-1. Scale bar = 50 μm. (k) After 14 days, mature neurons were immunostained for MAP2. Scale bar = 50 μm. (l,m) Quantification of Tuj-1 + immature neurons (l) and MAP2 + mature neurons (m) . ** P < 0.01, *** P < 0.001. (n,o) qPCR analysis of transcription factors associated with neuronal differentiation at 7 (n) and 14 (o) days. * P < 0.05, ** P < 0.01, *** P < 0.001. (p) Schematic of the NSPC axon growth assay. (q) After 7 days in proliferation culture, axons were immunostained for GAP43. Scale bar = 200 μm. (r) Quantification of axon length. (s) qPCR analysis of transcription factors regulating axon growth. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following primary antibodies were used: Nestin (rabbit, Proteintech, 1:400; NSPCs), SOX2 (rabbit, Proteintech, 1:400; NSPCs), PAX6 (rabbit, Proteintech, 1:400; NSPCs), TUJ1 (mouse, Proteintech, 1:400; immature neurons), MAP2 (mouse, Abcam, 1:400; mature neurons), GFAP (rat, Thermo Fisher, 1:400; astrocytes), GAP43 (rabbit, Proteintech, 1:400; axons), MMP-9 (rabbit, Abclonal, 1:400; secreted MMP-9), TNF-α (mouse, Proteintech, 1:400; pro-inflammatory cytokines), IL-10 (mouse, Proteintech, 1:400; anti-inflammatory cytokines), CD68 (rat, Abcam, 1:400; activated immune cells), CSPGs (mouse, Cell Signaling Technology, 1:400; glial scar), synaptophysin (SYP, rabbit, Proteintech, 1:400; presynaptic membranes), and cAMP (rabbit, Proteintech, 1:400; intracellular second messenger).

Techniques: In Vitro, Isolation, Differentiation Assay, Analysis, Growth Assay