Journal: Frontiers in Cell and Developmental Biology
Article Title: Cerebrospinal fluid-contacting neurons: a promising source for adult neural stem cell transplantation in spinal cord injury treatment
doi: 10.3389/fcell.2025.1549194
Figure Lengend Snippet: Transplanted CSF-CNs exhibit multipotential differentiation ability in vivo . (A) Representative images showing CSF-cNs (GFP + cells) co-labeled with the mature neuron marker NeuN. Co-labeling with NeuN was observed in CSF-cNs (GFP + cells) at 7 and 14 days post-transplantation. (B) Representative images showing CSF-cNs (GFP + cells) co-labeled with the oligodendrocyte marker O4. Co-labeling with O4 was detected in CSF-cNs (GFP + cells) at 7 and 14 days post-transplantation. (C) Representative images showing CSF-cNs (GFP + cells) co-labeled with the astrocyte marker S100β. Co-labeling with S100β was observed in CSF-cNs (GFP + cells) at 7 days post-transplantation. (D) Representative images showing transplanted CSF-cNs (GFP + cells) co-labeled with the immature neuron marker βIII tubulin. Co-labeling with βIII tubulin was detected in CSF-cNs (GFP + cells) at 3, 7, and 14 days post-transplantation. (E) Representative images showing CSF-cNs (GFP + cells) co-labeled with the GABA marker. Co-labeling with GABA was detected in CSF-cNs (GFP + cells) at 3, 7, and 14 days post-transplantation. White boxes indicate co-labeled positive cells; the enlarged sections on the right side of images (A–E) (a–e) show the positive cells within the boxes. (F) Quantitative analysis of the results from experiments (A–E) . By day 7 post-transplantation, CSF-cNs (GFP + cells) began differentiating into mature neurons and oligodendrocytes, with further increases observed by day 14. Conversely, differentiation into a small number of astrocytes was detected only at day 7 post-transplantation and had disappeared by day 14. DPI = days post-injury, SCI = spinal cord injury, N = 6.
Article Snippet: After deparaffinization, the tissue was permeabilized using a solution containing 0.25% Triton X-100, followed by blocking with a goat serum solution for 1 h. The primary antibodies used in this study included Pkd2l1 (AB9084, 1:700, Merck Millipore, MA, United States), Nestin (sc23927, 1:200, Santa Cruz Biotechnology, Dallas, TX, United States), Sox2 (1:400, CST, Danvers, MA, United States), GFAP (1:300, CST, Danvers, MA, United States), CD133 (66666-1-lg, 1:400, Proteintech, United States), EGFR (66455-1-lg, 1:500, Proteintech, United States), PCNA (10205-2-AP, 1:300, Proteintech, United States), NeuN (MAB377, 1:400, Millipore, United States), O4 (MAB1326, 1:400, RD, United States), β III tubulin (66240-1-Ig, 1:200, Proteintech, United States), S100β (66616-1-lg, 1:500, Proteintech, United States), and GABA (A2052, 1:200, Millipore, United States).
Techniques: In Vivo, Labeling, Marker, Transplantation Assay
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