Journal: Cell reports
Article Title: Adult leptomeningeal vestigial neural crest-derived multipotent cells promote vascular repair after stroke
doi: 10.1016/j.celrep.2025.116747
Figure Lengend Snippet: (A) In adult C57BL/6J mice, CD271 + cells (subset 5) were detected in the meninges, cranial sutures, and skull marrow but absent from the cortex. DAPI labels nuclei. (B and C) CD271 + cells isolated from leptomeninges-enriched tissue showed high expression of Tfap2 , Foxd3 , and Sox10 compared with cortical cells ( n = 3). All data are mean ± SD. (D and E) Human leptomeninges also contained CD271 + cells. After MACS isolation (D) and short-term expansion, these cells expressed HNK-1, Sox10, and CXCR4 (E). (F) CD271 + meningeal cells increased ipsilaterally after tMCAO or dMCAO and migrated into the injured cortex only on the infarcted side. (G) On day 3 after tMCAO, CD271 + cells upregulated Ngfr , Tfap2 , Foxd3 , Prrx2 , and Twist1 , while Sox10 was downregulated ( n = 4). All data are mean ± SD. (H–M) Endogenous meningeal NCC labeling and fate tracking. (H) AAV-CAG-tdTomato was applied to the leptomeninges; two-photon imaging was performed before and 3 days post tMCAO. (I) Flow cytometry confirmed that tdTomato + cells co-expressed CD271. (J and K) tdTomato + cells migrated from meninges into the ipsilateral cortex and localized near CD31 + vessels. (L) tdTomato + cells co-expressed CD271 on day 3. (M) A subset expressed NeuN by day 10, consistent with alternative NC-derived neuronal fates. (N–R) Transplantation of exogenous CD271 + NCCs to confirm meningeal-to-cortex migration. (N) CD271 + cells were isolated from injured cortex 3 days post tMCAO. (O) These cells formed vimentin + /Foxd3 + spheres. (P) Under defined conditions, they differentiated into NeuroTrace + neurons (NGF + brain-derived neurotrophic factor [BDNF]) or SMA + smooth muscle (TGF-β + fetal bovine serum [FBS]). (Q) PKH26-labeled CD271 + cells were transplanted onto the meninges of a second mouse on day 1 post tMCAO. (R) By day 3, PKH26 + cells localized adjacent to CD31 + vessels in cortical layers II/III and IV. All data are mean ± SD.
Article Snippet: TWIST1 , Applied Biosystems , Mm00442036_m1.
Techniques: Isolation, Expressing, Labeling, Imaging, Flow Cytometry, Derivative Assay, Transplantation Assay, Migration
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![(A) In adult C57BL/6J mice, CD271 + cells (subset 5) were detected in the meninges, cranial sutures, and skull marrow but absent from the cortex. DAPI labels nuclei. (B and C) CD271 + cells isolated from leptomeninges-enriched tissue showed high expression of Tfap2 , Foxd3 , and Sox10 compared with cortical cells ( n = 3). All data are mean ± SD. (D and E) Human leptomeninges also contained CD271 + cells. After MACS isolation (D) and short-term expansion, these cells expressed HNK-1, Sox10, and CXCR4 (E). (F) CD271 + meningeal cells increased ipsilaterally after tMCAO or dMCAO and migrated into the injured cortex only on the infarcted side. (G) On day 3 after tMCAO, CD271 + cells upregulated Ngfr , Tfap2 , Foxd3 , Prrx2 , and <t>Twist1</t> , while Sox10 was downregulated ( n = 4). All data are mean ± SD. (H–M) Endogenous meningeal NCC labeling and fate tracking. (H) AAV-CAG-tdTomato was applied to the leptomeninges; two-photon imaging was performed before and 3 days post tMCAO. (I) Flow cytometry confirmed that tdTomato + cells co-expressed CD271. (J and K) tdTomato + cells migrated from meninges into the ipsilateral cortex and localized near CD31 + vessels. (L) tdTomato + cells co-expressed CD271 on day 3. (M) A subset expressed NeuN by day 10, consistent with alternative NC-derived neuronal fates. (N–R) Transplantation of exogenous CD271 + NCCs to confirm meningeal-to-cortex migration. (N) CD271 + cells were isolated from injured cortex 3 days post tMCAO. (O) These cells formed vimentin + /Foxd3 + spheres. (P) Under defined conditions, they differentiated into NeuroTrace + neurons (NGF + brain-derived neurotrophic factor [BDNF]) or SMA + smooth muscle (TGF-β + fetal bovine serum [FBS]). (Q) PKH26-labeled CD271 + cells were transplanted onto the meninges of a second mouse on day 1 post tMCAO. (R) By day 3, PKH26 + cells localized adjacent to CD31 + vessels in cortical layers II/III and IV. All data are mean ± SD.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_9227/pmc12869227/pmc12869227__nihms-2142934-f0005.jpg)