Additional file 1 of Neural precursor cell delivery induces acute post-ischemic cerebroprotection, but fails to promote long-term stroke recovery in hyperlipidemic mice due to mechanisms that include pro-inflammatory responses associated with brain hemorrhages
Creators: name: Yin, Dongpei
name: Wang, Chen
name: Qi, Yachao
name: Wang, Ya-Chao
name: Hagemann, Nina
name: Mohamud Yusuf, Ayan
name: Dzyubenko, Egor
name: Kaltwasser, Britta
name: Tertel, Tobias
name: Giebel, Bernd
name: Gunzer, Matthias
name: Popa-Wagner, Aurel
name: Doeppner, Thorsten R.
name: Hermann, Dirk M.
Publisher: name: figshare
Year: 2023
Subjects: subject: Medicine
subject: Cell Biology
subject: Neuroscience
subject: Physiology
subject: FOS: Biological sciences
subject: Pharmacology
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Hematology

description: Additional file 1: Figure S1. Gating strategies used for analyzing brain and peripheral blood leukocytes and leukocyte activation via flow cytometry. In two sets of studies, lymphoid and myeloid cells were examined. Figure S2. NPC administration does not influence microvascular ICAM-1 expression, but hemorrhage incidence is higher in NPC-treated hyperlipidemic compared to normolipidemic mice. (A) Intercellular adhesion molecule-1 (ICAM-1) abundance on ischemic microvessels assessed by immunohistochemistry and (B) hemorrhage incidence evaluated by diaminobenzidine staining of normolipidemic mice on normal diet and hyperlipidemic mice on Western diet, which were exposed to 30 min intraluminal middle cerebral artery occlusion (MCAO) and intravenously treated with vehicle (200 µl of 0.1 M phosphate-buffered saline [PBS]) or adult NPCs (106 cells in 200 µl of 0.1 M PBS) immediately after reperfusion, followed by animal sacrifice at 48 h post-MCAO. Representative sections are shown. Data are medians (lines inside boxes)/means (crosses inside boxes) ± interquartile ranges with minimum/maximum values as whiskers. No significant group differences were noted (n = 12 mice for normal diet/vehicle, n = 12 for normal diet/NPC, n = 10 for Western diet/vehicle, n = 11 for Western diet/NPC). Scale bar, 50 μm. Figure S3. NPC administration does not influence neurological deficits in hyperlipidemic mice. Motor-coordination performance in (A) the Rotarod test and (B) the tight rope test, (C) body weight and (D) laser Doppler flow recordings above the core of the middle cerebral artery territory of mice on Western diet exposed to 30 min intraluminal MCAO, which were intravenously treated with vehicle or NPCs immediately after reperfusion (once only, dosing as before), followed by animal sacrifice after 56 days. Note that NPCs did not influence motor-coordination performance. For hyperlipidemic mice treated three times with NPCs see Fig. 6. Data are means ± S.D. values. No significant group differences were noted (n = 11 mice for Western diet/vehicle, n = 13 for Western diet/NPC). Figure S4. NPC delivery does not influence brain atrophy and glial scar formation in the chronic stroke phase in hyperlipidemic mice. (A) Brain atrophy, (B) striatal atrophy and (C) corpus callosum thickness assessed by cresyl violet staining, and (D) glial fibrillary acidic protein (GFAP) expression in the striatum assessed by immunohistochemistry in MCAO mice on Western diet, which were intravenously treated with vehicle or NPCs immediately after reperfusion (once only, dosing as before), followed by animal sacrifice after 56 days. For hyperlipidemic mice treated three times with NPCs see Fig. 7. Data are medians (lines inside boxes)/means (crosses inside boxes) ± interquartile ranges with minimum/maximum values as whiskers. No significant group differences were noted (n = 11 mice for Western diet/vehicle, n = 13 for Western diet/NPC). Scale bar, 1 mm [in (A–C)] and 50 μm [in (D)]. Table S1. Scoring of tight rope test performance. Table S2. Antibodies used for flow cytometry.