Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Brain pericytes increase glutamate uptake in astrocyte cultures. (A) Double immunofluorescence staining for CD13/EAAT1 (top left panel), CD13/EAAT2 (top right panel), GFAP/EAAT1 (bottom left panel), and GFAP/EAAT2 (bottom right panel) in intact mouse hippocampus sections. CD13 and GFAP signals appear green, while EAAT1 and EAAT2 signals appear red. (B) Regression analyses of cell/medium ratios of [ 3 H]-L-glutamate ([ 3 H]-L-Glu) uptake versus incubation time in astrocytes cultured without pericytes (astrocyte monocultures) or with pericytes (pericyte co-cultures). Astrocytes in monocultures and pericyte co-cultures were incubated in uptake buffer containing [ 3 H]-L-Glu for 1, 2, 5, and 10 min. A linear regression analysis was also performed to obtain the rate of [ 3 H]-L-Glu uptake. The slope of the lines was 9.48 in astrocyte monocultures ( r 2 = 0.90) and 13.54 in pericyte co-cultures ( r 2 = 0.89). Values are expressed as means ± SEM (n = 4 per point). * p < 0.05, between astrocyte monocultures and pericyte co-cultures.

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Double Immunofluorescence Staining, Incubation, Cell Culture

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Brain pericytes increase the Na + -dependent glutamate uptake in astrocytes. (A) Effect of Na + -free conditions on [ 3 H]-L-Glu uptake by astrocytes in monocultures or pericyte co-cultures. Results are expressed as the % of [ 3 H]-L-Glu uptake by astrocytes in monocultures incubated with uptake buffer containing Na + and [ 3 H]-L-Glu (Control; 113.96 ± 2.97 μL/mg protein). Values expressed are means ± SEM (n = 3); * p < 0.05, ** p < 0.01. (B) Effect of the EAAT1 inhibitor UCPH-101 (10 μM) on [ 3 H]-L-Glu uptake by astrocytes in monocultures or pericyte co-cultures. (C) Effect of the EAAT2 inhibitor DHK (500 μM) on [ 3 H]-L-Glu uptake by astrocytes in monocultures or pericyte co-cultures. Results are expressed as the % decrease from [ 3 H]-L-Glu uptake in astrocytes of the corresponding vehicle-treated group: 92.11 ± 12.34 μL/mg protein (B) and 75.32 ± 7.45 μL/mg protein (C) in astrocyte monocultures and 111.15 ± 14.40 μL/mg protein (B) and 98.42 ± 10.88 μL/mg protein (C) in pericyte co-cultures, respectively. Values expressed are the means ± SEM (n = 5–7); * p < 0.05.

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Incubation, Control

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Effect of pericytes on the protein expression levels of EAAT1 and EAAT2 in astrocytes. Representative western blot images (top panels) and densitometric analyses (bottom panels) of the expression of EAAT1 (A) and EAAT2 (B) in astrocytes. The protein levels of EAAT1 and EAAT2 were normalized to those of GAPDH. Results are expressed as the % of expression in astrocyte monocultures. Values expressed are the means ± SEM (n = 3).

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Expressing, Western Blot

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Treatment with pericyte-conditioned medium (pericyte-CM) upregulates glutamate uptake in astrocyte monocultures. Astrocyte monocultures exposed to culture medium (0 %), 50 % pericyte-CM, or 100 % pericyte-CM for 3 days were incubated with uptake buffer containing [ 3 H]-L-Glu for 10 min. Results are expressed as the % of [ 3 H]-L-Glu uptake by astrocytes in monocultures treated with 0 % pericyte-CM (72.35 ± 8.84 μL/mg protein). Values expressed are the means ± SEM (n = 3–6); ** p < 0.01.

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Incubation

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Brain pericytes increase glutamate uptake in astrocyte cultures. (A) Double immunofluorescence staining for CD13/EAAT1 (top left panel), CD13/EAAT2 (top right panel), GFAP/EAAT1 (bottom left panel), and GFAP/EAAT2 (bottom right panel) in intact mouse hippocampus sections. CD13 and GFAP signals appear green, while EAAT1 and EAAT2 signals appear red. (B) Regression analyses of cell/medium ratios of [ 3 H]-L-glutamate ([ 3 H]-L-Glu) uptake versus incubation time in astrocytes cultured without pericytes (astrocyte monocultures) or with pericytes (pericyte co-cultures). Astrocytes in monocultures and pericyte co-cultures were incubated in uptake buffer containing [ 3 H]-L-Glu for 1, 2, 5, and 10 min. A linear regression analysis was also performed to obtain the rate of [ 3 H]-L-Glu uptake. The slope of the lines was 9.48 in astrocyte monocultures ( r 2 = 0.90) and 13.54 in pericyte co-cultures ( r 2 = 0.89). Values are expressed as means ± SEM (n = 4 per point). * p < 0.05, between astrocyte monocultures and pericyte co-cultures.

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Double Immunofluorescence Staining, Incubation, Cell Culture

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Brain pericytes increase the Na + -dependent glutamate uptake in astrocytes. (A) Effect of Na + -free conditions on [ 3 H]-L-Glu uptake by astrocytes in monocultures or pericyte co-cultures. Results are expressed as the % of [ 3 H]-L-Glu uptake by astrocytes in monocultures incubated with uptake buffer containing Na + and [ 3 H]-L-Glu (Control; 113.96 ± 2.97 μL/mg protein). Values expressed are means ± SEM (n = 3); * p < 0.05, ** p < 0.01. (B) Effect of the EAAT1 inhibitor UCPH-101 (10 μM) on [ 3 H]-L-Glu uptake by astrocytes in monocultures or pericyte co-cultures. (C) Effect of the EAAT2 inhibitor DHK (500 μM) on [ 3 H]-L-Glu uptake by astrocytes in monocultures or pericyte co-cultures. Results are expressed as the % decrease from [ 3 H]-L-Glu uptake in astrocytes of the corresponding vehicle-treated group: 92.11 ± 12.34 μL/mg protein (B) and 75.32 ± 7.45 μL/mg protein (C) in astrocyte monocultures and 111.15 ± 14.40 μL/mg protein (B) and 98.42 ± 10.88 μL/mg protein (C) in pericyte co-cultures, respectively. Values expressed are the means ± SEM (n = 5–7); * p < 0.05.

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Incubation, Control

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Effect of pericytes on the protein expression levels of EAAT1 and EAAT2 in astrocytes. Representative western blot images (top panels) and densitometric analyses (bottom panels) of the expression of EAAT1 (A) and EAAT2 (B) in astrocytes. The protein levels of EAAT1 and EAAT2 were normalized to those of GAPDH. Results are expressed as the % of expression in astrocyte monocultures. Values expressed are the means ± SEM (n = 3).

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Expressing, Western Blot

Journal: IBRO Neuroscience Reports

Article Title: Brain pericytes upregulate glutamate uptake by astrocytes in vitro through sodium-dependent transporter

doi: 10.1016/j.ibneur.2025.05.017

Figure Lengend Snippet: Treatment with pericyte-conditioned medium (pericyte-CM) upregulates glutamate uptake in astrocyte monocultures. Astrocyte monocultures exposed to culture medium (0 %), 50 % pericyte-CM, or 100 % pericyte-CM for 3 days were incubated with uptake buffer containing [ 3 H]-L-Glu for 10 min. Results are expressed as the % of [ 3 H]-L-Glu uptake by astrocytes in monocultures treated with 0 % pericyte-CM (72.35 ± 8.84 μL/mg protein). Values expressed are the means ± SEM (n = 3–6); ** p < 0.01.

Article Snippet: Human brain-derived pericytes (#1200, Lot 27194; ScienCell Research Laboratories) and human brain-derived astrocytes (#CC-2565, Lot 0000647218; Lonza Bioscience) at passages 4–8 and 4–5, respectively, were used.

Techniques: Incubation

Journal: Neural Regeneration Research

Article Title: Advances in therapies using mesenchymal stem cells and their exosomes for treatment of peripheral nerve injury: state of the art and future perspectives

doi: 10.4103/NRR.NRR-D-24-00235

Figure Lengend Snippet: Typical examples of preclinical investigations on the application of various types of mesenchymal stem cells in peripheral nerve injury using different delivery methods

Article Snippet: One study showed that a model of PNI in Sprague–Dawley rats exhibited greater improvements in the restoration rate of gastrocnemius-muscle wet weight, sciatic function index (SFI), nerve conduction velocity (NCV), and myelin-sheath thickness after IM injection of bone marrow-derived MSCs (BMSCs), compared with IV delivery of BMSCs or phosphate-buffered solution (Wang et al., 2015a).

Techniques: Injection, IV Injection, Saline, Microinjection, Functional Assay, In Vitro, In Vivo, Transplantation Assay, Expressing

Journal: Neural Regeneration Research

Article Title: Advances in therapies using mesenchymal stem cells and their exosomes for treatment of peripheral nerve injury: state of the art and future perspectives

doi: 10.4103/NRR.NRR-D-24-00235

Figure Lengend Snippet: Historical milestones in preclinical investigations applying various types of MSCs in peripheral nerve injury. Created with BioRender.com. MSC: Mesenchymal stem cell.

Article Snippet: One study showed that a model of PNI in Sprague–Dawley rats exhibited greater improvements in the restoration rate of gastrocnemius-muscle wet weight, sciatic function index (SFI), nerve conduction velocity (NCV), and myelin-sheath thickness after IM injection of bone marrow-derived MSCs (BMSCs), compared with IV delivery of BMSCs or phosphate-buffered solution (Wang et al., 2015a).

Techniques:

Journal: Neural Regeneration Research

Article Title: Advances in therapies using mesenchymal stem cells and their exosomes for treatment of peripheral nerve injury: state of the art and future perspectives

doi: 10.4103/NRR.NRR-D-24-00235

Figure Lengend Snippet: Mechanisms underlying MSC therapy for peripheral nerve injury. (A) MSCs can differentiate into Schwann cell–like cells that produce growth factors, secrete neurotrophic factors, clear debris, and promote vascularization and myelination. (B) MSCs enhance nerve regeneration through their paracrine effects, which facilitate the transition of pro-inflammatory T helper 1 (Th1) cells to anti-inflammatory Th2 cells, recruit macrophages, promote M2-type polarization, and support vascularization. (C) Direct cell-to-cell contact between MSCs and other cells, such as pro-inflammatory macrophages, inhibits T-cell proliferation, and facilitates the transition of M1 macrophages to the M2 type, thereby alleviating excessive inflammation. Cell-to-cell contacts also provide approaches for the transport of molecules and organelles such as mitochondria between cells. (D) MSCs enhance the expression of transcription factors such as Krox-20/EGR2, which increase myelin proteins and promote myelination. Created with Adobe Illustrator TM 2019, with elements sourced from Servier Medical Art. Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licenses/by/3.0/). MSC: Mesenchymal stem cell.

Article Snippet: One study showed that a model of PNI in Sprague–Dawley rats exhibited greater improvements in the restoration rate of gastrocnemius-muscle wet weight, sciatic function index (SFI), nerve conduction velocity (NCV), and myelin-sheath thickness after IM injection of bone marrow-derived MSCs (BMSCs), compared with IV delivery of BMSCs or phosphate-buffered solution (Wang et al., 2015a).

Techniques: Expressing

Journal: Neural Regeneration Research

Article Title: Advances in therapies using mesenchymal stem cells and their exosomes for treatment of peripheral nerve injury: state of the art and future perspectives

doi: 10.4103/NRR.NRR-D-24-00235

Figure Lengend Snippet: Mechanisms of mesenchymal stem cells and their exosomes for treating peripheral nerve injury. Created using Adobe Illustrator™ 2019, with elements sourced from Servier Medical Art. Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licenses/by/3.0/). MSCs: Mesenchymal stem cells; M1: M1-polarized macrophage; M2: M2-polarized macrophage; Th1: T helper 1 cell; Th2: T helper 2 cell.

Article Snippet: One study showed that a model of PNI in Sprague–Dawley rats exhibited greater improvements in the restoration rate of gastrocnemius-muscle wet weight, sciatic function index (SFI), nerve conduction velocity (NCV), and myelin-sheath thickness after IM injection of bone marrow-derived MSCs (BMSCs), compared with IV delivery of BMSCs or phosphate-buffered solution (Wang et al., 2015a).

Techniques:

Journal: Neural Regeneration Research

Article Title: Human induced pluripotent stem cell–derived therapies for regeneration after central nervous system injury

doi: 10.4103/NRR.NRR-D-24-00901

Figure Lengend Snippet: Differentiation and identification of iPSC-derived cells. depicts the differentiation pathways of the cells derived from human iPSCs covered in this text for regenerative medicine. 1: Oikari et al. (2016); 2: Arber et al. (1999); 3: Phillis (2005); 4: Hu et al. (2010); 5: Hevner et al. (2001); 6: Forrest et al. (2023); 7: Zhou and Anderson (2002); 8: Zhu et al. (2008); 9: Seeker and Williams (2022); 10: Varga et al. (2022); 11: Molyneaux et al. (2007); 12: Zholudeva et al. (2024); 13: Glasgow et al. (2005); 14: Falgairolle and O’Donovan (2019); 15: Thaler et al. (2002); 16: Duan et al. (2016); 17: Janke and Magiera (2020); 18: Babkina et al. (2024); 19: Ruan and Elyaman (2022); 20: Schwabenland et al. (2021); 21: Bernal and Arranz (2018); 22: Thiry et al. (2020). Created with BioRender.com. Aldh1l1: Aldehyde dehydrogenase 1 family member L1; Aqp4: aquaporin 4; ChAT: choline acetyltransferase; Ctip2: COUP-TF interacting protein 2; DCX: doublecortin; Emx1/2: empty spiracles homeobox 1/2; Fezf2: forebrain embryonic zinc finger-like protein 2; Foxg1: forkhead box G1; GFAP: glial fibrillary acidic protein; GLT1: glutamate transporter 1; HB9 (MNX1): homeobox 1 (motor neuron and pancreas homeobox 1); Iba1: ionized calcium-binding adapter molecule 1; iPSCs: induced pluripotent stem cells; ISL1: insulin gene enhancer protein 1; Lhx3: LIM homeobox 3; MBP: myelin basic protein; MSI1: musashi RNA binding protein 1; Ngn2: neurogenin 2; NSE: neuron-specific enolase; Olig1/2: oligodendrocyte transcription factor 1/2; Pax6: paired box 6; PDGFRA: platelet-derived growth factor receptor alpha; Ptf1a: pancreas transcription factor 1a; Sox1: SRY-box transcription factor 1; Tbr1: T-box brain transcription factor 1; Tmem119: transmembrane protein 119; TUBB3: β-3-tubulin; VSX1/2: visual system homeobox 1/2.

Article Snippet: Lu et al., 2014: Long-distance axonal growth from human induced pluripotent stem cells after spinal cord injury , LV-GFP hiPSC- derived NSCs Nestin +, Sox2+, SSEA4-, TRA-1- 81+ , Adult female athymic nude Rats (180-200 g, T-cell deficient, Harlan Laboratories) , C5 hemi- contusion SCI , Microinjection into lesion. Injected in Fibrin and Growth Factor Cocktail , 14 d post-SCI , 1.25×10 6 /5 μL , Successful differentiation, survival, and integration of the graft into the host. VGlut1 + , ChAT + , and 5HT + neurons from the graft identified. Transplanted cells found in various areas of the brain 3 months following transplant..

Techniques: Derivative Assay, Binding Assay, RNA Binding Assay

Journal: Neural Regeneration Research

Article Title: Human induced pluripotent stem cell–derived therapies for regeneration after central nervous system injury

doi: 10.4103/NRR.NRR-D-24-00901

Figure Lengend Snippet: Summary of transplant studies

Article Snippet: Lu et al., 2014: Long-distance axonal growth from human induced pluripotent stem cells after spinal cord injury , LV-GFP hiPSC- derived NSCs Nestin +, Sox2+, SSEA4-, TRA-1- 81+ , Adult female athymic nude Rats (180-200 g, T-cell deficient, Harlan Laboratories) , C5 hemi- contusion SCI , Microinjection into lesion. Injected in Fibrin and Growth Factor Cocktail , 14 d post-SCI , 1.25×10 6 /5 μL , Successful differentiation, survival, and integration of the graft into the host. VGlut1 + , ChAT + , and 5HT + neurons from the graft identified. Transplanted cells found in various areas of the brain 3 months following transplant..

Techniques: Transplantation Assay, Animal Model, Cell Counting, Virus, Isolation, Injection, Derivative Assay, Microinjection, Functional Assay, Migration, Expressing, Knock-In, Activity Assay, Control, Optogenetics, Inhibition, Saline, Shear, Encapsulation, Anterograde Tracing, In Vivo, Patch Clamp, In Vitro, Staining