Journal: Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie

Article Title: Bone Marrow Mesenchymal Stem Cell Administration Significantly Improves Outcome after Retinal Ischemia in Rats

doi: 10.1007/s00417-017-3690-1

Figure Lengend Snippet: Stimulus-Intensity responses for the a-wave (A), b-wave (B), P2 (C), sum of the root mean squared OP waves (OPRMS) (D) from electroretinograms in rats subjected to retinal ischemia for 55 min and the eyes injected 24 h later with BMSCs or control (PBS). The recovery was measured 7 days later; N= 8–9. Normalized ERG recovery data (Y-axis, see Methods) for the waves over a range of up to 10 flash intensities (x-axis) are shown. For the a-wave, data collection began at a higher initial flash intensity due to artifact and low amplitude at the two lower intensities. There was significant improvement with injection of BMSCs 24 h after ischemia on all of the ERG waves, although results were not statistically significant at all intensity levels. Data are shown as mean ± SEM. * P < 0.05 vs PBS vehicle control. Representative ERG traces for a- and b-waves, OP, and P2 from our lab are available in previous publications [16].

Article Snippet: To track the fate of BMSCs, we used rat green fluorescent-labeled BMSCs (GFP-BMSCs, Cyagen, Santa Clara, CA).

Techniques: Injection, Control

Journal: Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie

Article Title: Bone Marrow Mesenchymal Stem Cell Administration Significantly Improves Outcome after Retinal Ischemia in Rats

doi: 10.1007/s00417-017-3690-1

Figure Lengend Snippet: (A&C) Stimulus-Intensity responses for the STR (scotopic threshold response), for the positive STR (pSTR) and negative STR (nSTR) in non-ischemic eyes of rats subjected to retinal ischemia for 55 min and the eyes injected 24 h later with BMSCs or control (PBS). (B&D) STRs recorded from the ischemic eyes at baseline and after ischemia. The recovery was measured 7 days later. Absolute amplitude appears on the Y-axis (see Methods) and the 6 flash intensities are on the X-axis. There was significant improvement in the nSTR, but not the pSTR with injection of BMSCs 24 h after ischemia. Data are shown as mean + SEM. * P < 0.05 vehicle Day 7 vs baseline; # = P < 0.05 BMSC Day 7 vs baseline; x = P < 0.05 vehicle baseline vs BMSC baseline;. = P < 0.05 vehicle Day 7 vs BMSC day 7. N = 6. (E) Representative STR traces at the highest flash intensity, with BMSC-ischemic eye on top, and PBS-ischemic eye below

Article Snippet: To track the fate of BMSCs, we used rat green fluorescent-labeled BMSCs (GFP-BMSCs, Cyagen, Santa Clara, CA).

Techniques: Injection, Control

Journal: Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie

Article Title: Bone Marrow Mesenchymal Stem Cell Administration Significantly Improves Outcome after Retinal Ischemia in Rats

doi: 10.1007/s00417-017-3690-1

Figure Lengend Snippet: (A, C, G, E) Stimulus-Intensity responses for the a wave, b-wave, P2, and OPRMS in non-ischemic eyes of rats subjected to retinal ischemia for 55 min and the eyes injected 24 h later with BMSCs or control (PBS). (B, D, F, H) Stimulus intensity responses for a, b, P2, and OPRMS waves recorded from the ischemic eyes at baseline and after ischemia. The recovery was measured 7 days later. Absolute amplitude appears on the Y-axis (see Methods) and the flash intensities are on the X-axis. Data are shown as mean + SEM. There were no differences between non-ischemic values for the OPRMS (Fig 3E), and few for the a-wave. For the b-wave and P2, both injection of BMSCs and PBS vehicle decreased amplitudes of the waveforms, and the vehicle-injected eyes had generally higher baseline amplitudes compared to the BMSC-injected. There was a greater decrease in amplitudes of the waveforms from baseline to day 7 in ischemic eyes for all waveforms for the vehicle compared to the BMSC injected eyes. * P < 0.05 vehicle Day 7 vs baseline; # = P < 0.05 BMSC Day 7 vs baseline; x = P < 0.05 vehicle baseline vs BMSC baseline;. = P < 0.05 vehicle Day 7 vs BMSC day 7. N = 8–9

Article Snippet: To track the fate of BMSCs, we used rat green fluorescent-labeled BMSCs (GFP-BMSCs, Cyagen, Santa Clara, CA).

Techniques: Injection, Control

Journal: Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie

Article Title: Bone Marrow Mesenchymal Stem Cell Administration Significantly Improves Outcome after Retinal Ischemia in Rats

doi: 10.1007/s00417-017-3690-1

Figure Lengend Snippet: Ischemic and normal eyes were injected with GFP-BMSC 24 h post ischemia. In vivo imaging of the eyes was performed at 1, 7 and 21 days’ post injection (A). The upper and lower panels exhibit representative images of normal and ischemic eyes injected with BMSCs. (B) Representative flat mount images of BMSCs injected retina illustrate the penetration and incorporation of significantly increased number of GFP-BMSCs into the ischemic retinal tissue 7 days’ post injection compared to normal retina. DAPI was used to stain nuclei.

Article Snippet: To track the fate of BMSCs, we used rat green fluorescent-labeled BMSCs (GFP-BMSCs, Cyagen, Santa Clara, CA).

Techniques: Injection, In Vivo Imaging, Staining

Journal: Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie

Article Title: Bone Marrow Mesenchymal Stem Cell Administration Significantly Improves Outcome after Retinal Ischemia in Rats

doi: 10.1007/s00417-017-3690-1

Figure Lengend Snippet: Fluorescent TUNEL staining for evaluation of apoptosis in 10-μm thick retinal cryosections: BMSC injection into the retina significantly reduced apoptotic cell death in the ischemic retinae 24 h post injection (B) as compared to PBS injected ischemic control (A). White arrows denote TUNEL positive cells (green) co-localized with DAPI (blue). Brackets denote the retinal ganglion cell layer. Magnification = 40X. Scale bar = 15 μm. (C) Retinal ganglion cells undergoing apoptosis in PBS injected control ischemic retina identified by co-localization of TUNEL and Thy1 (retinal ganglion cell marker, red). Magnification = 40X. Scale bars = 20 μm. (D) Representative Western blot images for cleaved caspase-3 in retinal lysates obtained from PBS or BMSC ischemic injected eyes and paired normal eyes, at 24 h after injection of PBS or BMSCs. Levels of cleaved caspase-3 were normalized to β-arrestin. Data are shown as mean ± SEM. * indicates P < 0.05 for ischemic compared to the paired normal retina within a group (PBS or BMSC); # indicates P < 0.05 between the ischemic retinae of PBS and BMSC. N = normal, non-ischemic retinae, and I = ischemic retinae. N = 5 per group.

Article Snippet: To track the fate of BMSCs, we used rat green fluorescent-labeled BMSCs (GFP-BMSCs, Cyagen, Santa Clara, CA).

Techniques: TUNEL Assay, Staining, Injection, Control, Marker, Western Blot

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Animal experimental flow chart. Exo miR-23b : exosomes derived from BMSCs transfected with miR-23b; Exo miR-NC : exosomes derived from BMSCs transfected with miR-NC; GSH: reduced glutathione; GSSG: oxidized glutathione disulfide; HE: hematoxylin-eosin; ICH: intracerebral hemorrhage; MDA: malondialdehyde; PBS: phosphate-buffered saline; ROS: reactive oxygen species; RT-qPCR: reverse transcription quantitative polymerase chain reaction; SOD: superoxide dismutase; TUNEL: terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: Derivative Assay, Transfection, Saline, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction, TUNEL Assay, End Labeling

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Identification of exosomes derived from BMSCs overexpressing miR-23b. (A) MiR-23b expression in BMSCs detected by RT-qPCR after lentivirus transfection. (B) Transmission electron microscope scanning images of exosomes. The exosomes derived from the BMSCs showed a round cup-shaped and complete structure. Scale bars: 100 nm. (C) Western blot bands detecting exosome markers CD63, TSG101, and CD81. (D) MiR-23b expression levels in different exosome groups detected by RT-qPCR. Data are shown as the mean ± SD ( n = 3). * P < 0.05 (Student’s t -test). BMSCs: Bone marrow mesenchymal stem cells; Exo miR-23b : exosomes derived from BMSCs transfected with miR-23b; Exo miR-NC : exosomes derived from BMSCs transfected with miR-NC; RT-qPCR: reverse transcription quantitative polymerase chain reaction.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Transfection, Transmission Assay, Microscopy, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Exosomal miR-23b inhibits oxidative stress in microglia BV2 cells in vitro . (A) miR-23b levels in microglia BV2 cells after administration of different exosome groups under hemin stimulation. (B) Representative images of ROS staining. The hemin group showed more ROS staining than that in the PBS group, and antioxidant NAC administration further reduced the ROS staining compared with that in the hemin group. ROS levels were decreased in both exomiR-23b and exomiR-NC groups compared with those in the hemin group, and the exomiR-23b group showed less ROS staining than that in the exomiR-NC group. Scale bars: 50 μm. (C) Quantification of relative fluorescence intensity of ROS. (D) MDA levels were measured by economical kits. (E) SOD levels were measured by economical kits. (F) The GSH/ GSSG ration was calculated on the basis the GSH and GSSG levels which evaluated by economical kits. Data are shown as the mean ± SD ( n = 6). $ P < 0.05, $$ P < 0.01, vs . PBS group; * P < 0.05, ** P < 0.01, vs . hemin model group; # P < 0.05, vs . exomiR-NC group (one-way analysis of variance followed by Newman-Keuls post hoc analysis). PBS: Control; PBS + NAC: treated with 1 mM N-acetylcysteine; Hemin: treated with 60 μM hemin for 24 hours; Hemin + NAC: treated with 1 mM N-acetylcysteine 2 hours after hemin stimulation; exomiR-NC group: treated with miR-NC transfected BMSCs-exosomes after hemin stimulation; exomiR-23b group: treated with miR-23b transfected BMSCs-exosomes after hemin stimulation. BMSC: Bone marrow mesenchymal stem cell; GSH: reduced glutathione; GSSG: oxidized glutathione disulfide; ICH: intracerebral hemorrhage; MDA: malondialdehyde; NAC: N-acetylcysteine; PBS: phosphate-buffered saline; ROS: reactive oxygen species; SOD: superoxide dismutase.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: In Vitro, Staining, Fluorescence, Control, Transfection, Saline

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Exosomal miR-23b alleviates NLRP3 inflammasome-mediated pyroptosis in microglia BV2 cells and protects neuronal cells in vitro . (A, B) Expression and data analysis of pyroptosis-related proteins in different groups of microglia BV2 cells. (C, D) IL-1β (C) and IL-18 (D) levels in microglia BV2 cells were evaluated by ELISA. (E) Cell viabilities of hippocampal neuronal HT22 cells by CCK8 tests. (F) Cell death rates of HT22 cells by LDH assays. Data are shown as the mean ± SD ( n = 6). $$ P < 0.01, vs . PBS group; * P < 0.05, ** P < 0.01, vs . hemin group; # P < 0.05, ## P < 0.01, vs . exo miR-NC group (one-way analysis of variance followed by Newman-Keuls post hoc analysis). PBS group: Control; Hemin group: treated with 60 μM hemin for 24 hours; exo miR-NC group: treated with miR-NC transfected BMSCs-exosomes after hemin stimulation; exo miR-23b group: treated with miR-23b transfected BMSC-exosomes after hemin stimulation. BMSC: Bone marrow mesenchymal stem cell; CCK8: cell counting kit-8; ELISA: enzyme linked immunosorbent assay; GSDMD-N: N-terminal fragment of gasdermin D; IL-18: interleukin-18; IL-1β: interleukin-1β; LDH: lactate dehydrogenase; NLRP3: NOD-like receptor family pyrin domain containing 3; PBS: phosphate-buffered saline.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: In Vitro, Expressing, Enzyme-linked Immunosorbent Assay, Control, Transfection, Cell Counting, Saline

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: BMSC-exosomal miR-23b attenuates oxidative stress by regulating the PTEN/Nrf2 pathway in microglia BV2 cells in vitro . (A) Western blot bands for PTEN, Nrf2 (cytoplasmic and nuclear), and HO-1 proteins in microglia BV2 cells from different groups. (B) Quantitative analysis of PTEN, Nrf2 (cytoplasmic and nuclear), and HO-1. (C) MDA levels were measured by economical kits. (D) SOD levels were measured by economical kits. (E) GSH/GSSG ratio was calculated on the basis of the GSH and GSSG levels which evaluated by economical kits. Data are shown as the mean ± SD ( n = 6). * P < 0.05, ** P < 0.01, vs . hemin group; # P < 0.05, vs . exo miR-23b group (one-way analysis of variance followed by Newman-Keuls post hoc analysis). Hemin: Treated with 60 μM hemin for 24 hours; exo miR-23b group: treated with miR-23b transfected BMSCs-exosomes after hemin stimulation; exo miR-23b + PTEN: treated with pcDNA3.1-PTEN plasmids and miR-23b transfected BMSCs-exosomes after hemin stimulation. BMSC: Bone marrow mesenchymal stem cell; GSH: reduced glutathione; GSSG: oxidized glutathione disulfide; HO-1: heme oxygenase-1; MDA: malondialdehyde; Nrf2: nuclear factor erythroid-2-related factor 2; PTEN: phosphatase and tensin homolog deleted on chromosome 10; SOD: superoxide dismutase.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: In Vitro, Western Blot, Transfection