Journal: Frontiers in Cellular Neuroscience

Article Title: Hypoxic culture of umbilical cord mesenchymal stem cell-derived sEVs prompts peripheral nerve injury repair

doi: 10.3389/fncel.2022.897224

Figure Lengend Snippet: Cellular localization and uptake of fluorescently labeled sEVs with SCs. (A) Cellular colocalization: SP8 was used to photograph sEVs prelabeled with PKH67 (green fluorescence) with Hoechst 33,342 (blue fluorescence)-stained SC nuclei. (B) Western blot: Detection of ERK1/2, ZEB2, and c-JUN expression levels in NC Schwann cells and after 48-h treatment with hypoxia sEVs. (C) Real-time PCR: Inflammatory and other restoration-related factors (IL-1β, IL-6, TNF-α, etc.) were detected. (D) ELISA test: GDNF NDF and NT-3 in the supernatant of Schwann cell preparation. Statistical significance, * p < 0.05,** p < 0.01, and **** p < 0.0001. *Significant difference.

Article Snippet: The following ELISA kits were: mouse glial cell line-derived neurotrophic factor (GDNF), ELISA kit (CSB-E07341m, CUSABIO), mouse neurotrophin 3 (NT-3), ELISA kit (CSB-E04687m, CUSABIO), and mouse nerve growth factor (NGF), ELISA kit (CSB-E04684m, CUSABIO).

Techniques: Labeling, Fluorescence, Staining, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

doi: 10.18632/oncotarget.12720

Figure Lengend Snippet: A. Light field of SSCs cultured with melatonin and GDNF, bar=50 μm. B. Cell density after being cultured with different cell mediums at 48 h. The initial number was 5*10 4 . C. QRT-PCR and western blot analysis of proliferation, self-renewal and Sertoli cell markers. D. Western blot analysis of proliferation and Sertoli cell markers. *, P<0.05,**, P<0.01.

Article Snippet: GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER).

Techniques: Cell Culture, Quantitative RT-PCR, Western Blot

Journal: Oncotarget

Article Title: Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

doi: 10.18632/oncotarget.12720

Figure Lengend Snippet: A. ELISA analysis of GDNF levels in the SSCs medium. B. Western Blot analysis of phosphorylation levels of AKT and ERK. *, P<0.05,**, P<0.01.

Article Snippet: GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Phospho-proteomics

Journal: Bioactive Materials

Article Title: Acellular nerve xenografts based on supercritical extraction technology for repairing long-distance sciatic nerve defects in rats

doi: 10.1016/j.bioactmat.2022.03.014

Figure Lengend Snippet: Evaluation of the cytotoxicity of the ANXs scaffold in vitro. (A, D) Living/dead double staining of Schwann cells grown on the ANXs scaffold for 3 days and 7 days (live: green, dead: red). (B, E) SEM images of Schwann cells growing on CD SD and CD + scCO 2 NG scaffolds for 7 days (the picture on the right is an enlarged view of the yellow area in the picture on the left). (C, F) Immunofluorescence images of Schwann cells growing on CD SD and CD + scCO 2 NG scaffolds for 7 days, respectively (S100: red, nucleus: blue). (G) Quantification of the number of live/dead double-stained Schwann cells in each region (0.36 mm 2 ). Data are presented as the mean ± SD (n = 3). (H) The CCK-8 assay was performed after 1, 3, 5 and 7 days of cell culture. Data are presented as the mean ± SD (n = 5). (I, J) Quantitative analysis of the GDNF and NGF expression levels of Schwann cells on the ANXs scaffold. Data are presented as the mean ± SD (n = 5). Statistical analysis: n.s. no significances, **p < 0.01, *p < 0.05.

Article Snippet: In brief, the medium of each group was centrifuged at 1500 rpm and 4 °C for 10 min, the concentration of NGF and BDNF in the supernatant was assessed using ELISA kits, the rat GDNF ELISA kit (EK0363, BOSTER, China) and the rat NGF/NGFβ ELISA kit (EK0471, BOSTER, China), and the absorbance of each well at 450 nm was determined using a spectrophotometer (EPOCH TAKE 3, Bio-Tek, USA).

Techniques: In Vitro, Double Staining, Immunofluorescence, Staining, CCK-8 Assay, Cell Culture, Expressing

Journal: Stem cells translational medicine

Article Title: Multimodal Delivery of Isogenic Mesenchymal Stem Cells Yields Synergistic Protection from Retinal Degeneration and Vision Loss.

doi: 10.5966/sctm.2016-0181

Figure Lengend Snippet: Figure 6. Mesenchymal stem cell (MSC)-mediated trophic support of visual function in vivo and retinal pigment epithelium (RPE) phagocytosis capacity ex vivo. (A): ERG b-wave amplitudes were higher in eyes treated with 353 MSC-CdMSRI (normalized to Contra. Eyes; n = 3), compared with BSSSRI (n = 5) at postnatal day 60 (p , .05). (B): ERG traces show higher b-wave amplitude from CdMSRI (66.8 mV) versus BSSSRI (24.6 mV). (C,D):Metabolic viability inRoyal CollegeofSurgeons RPE increased 24hoursafter exposureto 53CdM, which wasabrogated by preincubating CdM with blocking antibodies to select trophic factors (C) or by preincubating RPE cultures with antibodies to trophic factor receptors (D). CdM enhanced the total POSBound (E–H, quantified in I). (J, K): POSINT colocalization with phagolysosome marker, cathepsin-D, in BM (J, colocalized area in J’) was increased by CdM exposure (K, colocalized area in K’). (L): The peptides in (C) were screened for possible contribution to POSBound. (M):TotalPOSBoundwasabrogatedwithBDNFneutralization(82%65%ofIgG1control;*,p,.05)andwasrestoredwiththeadditionof50ng/ml BDNF to BM (197% 6 25% of IgG1 control; †, p , .05). Abbreviations: BDNF, brain-derived neurotrophic factor; bFGF, basic fibroblast growth factor;BM, basal medium; BSS, balanced salt solution; CdM,MSC-conditioned medium; CNTF, ciliary neutrophoric factor; CNTFR, CNTF receptor; Contra., contralateral (untreated); CTGF, connective tissue growth factor; CXCL12, C-X-C chemokine ligand 12; CXCR4, C-X-C chemokine receptor 4; DAPI, 49,6-diamidino-2-phenylindole; ERG, electroretinography; GDNF, glial-derived neurotrophic factor; GDNFR, GDNF receptor; GM, growth medium; IgG, immunoglobulin G; IgG1, immunoglobulin G1; POSBOUND, photoreceptor outer segments bound to RPE; POS, photoreceptor outer segments; POSINT, RPE-internalized POS; rBDNF, brain-derived neurotrophic factor receptor; SRI, subretinal injection; TrkA, TrkB, TrkC, tropomy- osin receptor kinase A, B, C; VEGF, vascular endothelial growth factor.

Article Snippet: For trophic factor and receptor blocking/neutralization studies, the following antibodies were used at 5 mg/ml: mouse immunoglobulin (Ig) G1 isotype control, brain-derived neurotrophic factor (BDNF) neutralizing (Thermo Fisher), fibroblast growth factor (FGF)-2, vascular endothelial growth factor (VEGF)-A, CXCL12, glial-derived neurotrophic factor (GDNF), BDNF, ciliary neurotrophic factor (CNTF) p75 nerve growth factor receptor (Santa Cruz Biotechnology Inc., Dallas, TX, http:// www.scbt.com), connective tissue growth factor (CTGF; Bioss Antibodies Inc., Woburn, MA, http://www. biossusa.com), nerve growth factor (NGF)/proNGF, tropomyosin receptor kinase (Trk)A, TrkB, TrkC, glial cell-derived neurotrophic factor receptor a1, rat p75NTR (Alomone Labs, Inc., Jerusalem, Israel, http://www.alomone.com).

Techniques: In Vivo, Ex Vivo, Blocking Assay, Marker, Control, Derivative Assay, Injection

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: Conditioned medium (CM) from cultures of oxygen-glucose deprivation (OGD) activated BV2 and microglia (MG) stimulates glial cell-derived neurotrophic factor (GDNF) production by bone marrow mesenchymal stem cells (BMSCs). (A) BV2 or (B) MG cells were exposed to OGD for 4 h before the addition of CM to BMSCs. After 24 h, GDNF concentrations in the BMSC culture supernatants were determined by ELISA. ** P < 0.01. *** P < 0.001.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: Tumor necrosis factor-α (TNFα), rather than interleukin-6 (IL6) or interleukin-1β (IL1β), elevates GDNF production by BMSCs. In (A) BV2 and (B) MG, OGD stimulates TNFα, IL6 and IL1β production. (C) GDNF production by BMSCs is promoted after treatment with TNFα for 24 h. (D) IL6 and (E) IL1β treatment for 24 h does not significantly change GDNF production by BMSCs. *** P < 0.001.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: GDNF repairs OGD-induced neuronal injury. (A) Neuronal viability is promoted by pretreatment with GDNF after OGD, and is inhibited by GDNF-specific siRNA transfection. (B) TNFα, IL6 or IL1β suppresses neuronal viability. * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Transfection

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: GDNF prevents OGD-induced neuronal injury. (A) Neuronal viability is promoted by pretreatment with GDNF before OGD, but is suppressed by anti-RET antibody treatment. (B) Neuronal apoptosis is reduced by GDNF pretreatment before OGD, but is improved by anti-RET antibody treatment. * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: GDNF regulates the mitogen-activated protein kinase (MAPK) kinase (MEK)/ERK and phosphoinositide-3-kinase (PI3K)/v-akt murine thymoma viral oncogene homolog (AKT) signaling pathways, but not the JNK/c-JUN pathway. Actin is used as an internal control. (A) The phosphorylated forms of MEK1/2, ERK1/2, PI3K and AKT (p-MEK1/2, p-ERK1/2, p-PI3K and p-AKT) are promoted by GDNF and suppressed by anti-RET antibody treatment. The phosphorylated forms of JNK and c-JUN (p-JNK or p-c-JUN) are barely changed by GDNF or anti-RET antibody treatment. (B) Relative protein levels based on Western blot results. NS, not significant. * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Protein-Protein interactions, Control, Western Blot

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: GDNF helps to maintain mitochondrial membrane potential (MMP) and inhibit neuronal injury induced by OGD. (A) Western blot showing GDNF up-regulates B cell lymphoma 2 (BCL2) and heat shock 60 kDa protein 1 (HSP60) protein levels in neurons after OGD. Actin is used as an internal control. (B) Relative BCL2 protein level based on Western blot results. (C) Relative HSP60 protein level based on Western blot results. (D) Lactate dehydrogenase (LDH) assay indicates that GDNF inhibits neuronal LDH leakage after OGD. (E) MMP assay indicates that GDNF helps to maintain neuronal MMP after OGD, but TNFα decreases MMP. NS, not significant. * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Membrane, Western Blot, Control, Lactate Dehydrogenase Assay, Mmp Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Activated Microglia Induce Bone Marrow Mesenchymal Stem Cells to Produce Glial Cell-Derived Neurotrophic Factor and Protect Neurons Against Oxygen-Glucose Deprivation Injury

doi: 10.3389/fncel.2016.00283

Figure Lengend Snippet: Schematic diagram of the effects of MG and BMSC on neurons revealed in this study. Following the stimulation by OGD, MG secretes TNFα, which further induces BMSCs to produce more GDNF. GDNF protects neurons against OGD-induced injury. Furthermore, TNFα secreted from MG induces inflammatory responses to suppress OGD-induced neuronal injury.

Article Snippet: GDNF concentrations in culture supernatants were determined using the rat GDNF ELISA Kit (Cusabio, Wuhan, China) according to the manufacturer’s instructions.

Techniques: