Journal: Scientific Reports

Article Title: Human Bone Marrow Endothelial Progenitor Cell Transplantation into Symptomatic ALS Mice Delays Disease Progression and Increases Motor Neuron Survival by Repairing Blood-Spinal Cord Barrier

doi: 10.1038/s41598-019-41747-4

Figure Lengend Snippet: Characteristics of disease outcomes in G93A mice receiving hBMEPCs at symptomatic stage. Transplanted ALS mice with cell dose of 1 × 10 6 at 4 weeks post-treatment ( A ) significantly maintained body weight, ( B ) better extended hindlimbs, ( C ) delayed loss in muscle strength, and ( D ) stayed longer on rotarod vs. media-injected mice. Notable beneficial effects on motor function were determined in G93A mice 2–3 weeks after cell transplantation. * p < 0.05, ** p < 0.01.

Article Snippet: Cryopreserved human bone marrow-derived endothelial progenitor cells (hBMEPCs) were purchased from CELPROGEN (Torrance, CA, USA).

Techniques: Injection, Transplantation Assay

Journal: Scientific Reports

Article Title: Human Bone Marrow Endothelial Progenitor Cell Transplantation into Symptomatic ALS Mice Delays Disease Progression and Increases Motor Neuron Survival by Repairing Blood-Spinal Cord Barrier

doi: 10.1038/s41598-019-41747-4

Figure Lengend Snippet: Immunocytochemical analysis of transplanted hBMEPCs in blood smears. For identification of transplanted hBMEPCs within blood circulation of treated mice, immunofluorescent staining with the human anti-vWF was performed in blood smears. ( A ) hBMEPCs were positive for vWF immunoexpression (red, arrowheads). A few cells did not express vWF (asterisk) likely due to damaging during smear preparation. ( B ) There was no detection of human cells by vWF marker in blood smears from media-treated mice (asterisks). ( C , C’) In blood smears from cell-treated ALS mice, some cells were identified with human vWF (red, arrowheads). Cells negative for vWF are noted by asterisks. The nuclei in all images are shown with DAPI. Scale bar in A is 20 µm and B-C’ is 50 µm.

Article Snippet: Cryopreserved human bone marrow-derived endothelial progenitor cells (hBMEPCs) were purchased from CELPROGEN (Torrance, CA, USA).

Techniques: Staining, Marker

Journal: bioRxiv

Article Title: BoneMA – Synthesis and Characterization of a Methacrylated Bone-derived Hydrogel for Bioprinting of Vascularized Tissues

doi: 10.1101/2020.03.02.974063

Figure Lengend Snippet: Representative fluorescence microscopy image demonstrating the range of patterns and geometries achieved by printing HMSC-laden BoneMA with a DLP printer (Ember). The cells were stained with F-Actin (green) after printing to enable confirmation of the shape and structure of the printed constructs through fluorescence microscopy. (Scale – 750 μm)

Article Snippet: To that end, human dental pulp stem cells (HDPSC) and human mesenchymal stem cells (HMSC) were used to assess cytocompatibility and bioprintability, while green fluorescent protein (GFP)-expressing human umbilical vein endothelial cells (HUVECs) (cAP0001GFP, Angio-proteomie, USA) were employed to investigate the vasculogenic potential of the synthesized biomaterial.

Techniques: Fluorescence, Microscopy, Staining, Construct

Journal: Research

Article Title: Fut2 Deficiency Promotes Intestinal Stem Cell Aging by Damaging Mitochondrial Functions via Down-Regulating α1,2-Fucosylation of Asah2 and Npc1

doi: 10.34133/research.0343

Figure Lengend Snippet: Fut2 deficiency in ISCs resulted in impairment of α1,2-fucosylation of mitochondrial-function-related proteins. (A) Volcano plot of differently expressed N-glycosylated proteins and sites in Fut2 knockout ISCs compared to WT. (B) Fold change of protein level and N-glycosylation level of mitochondrial-function-related proteins Asah2, Npc1, and Bsg. (C) IHC analysis of Asah2 in ileal sections of WT and Fut2 ΔISC mice. Scale bar, 100 μm. (D) The protein level of Npc1, Asah2, and Bsg in whole-cell lysate and UEA-I-enriched proteins of WT and Fut2 ΔISC ISCs. (E) Protein expression of mTOR and p-mTOR in WT and Fut2 ΔISC crypts. * P < 0.05, ** P < 0.01, and *** P < 0.001. ns, no significant.

Article Snippet: Ten micrograms of Asah2 recombinant protein (MedChemExpress, HY-P76735) and 250 nM mTOR inhibitor Torin1 (MedChemExpress, HY-13003) were used to treat organoids to confirm the role of Asah2 and Npc1 fucosylation.

Techniques: Knock-Out, Glycoproteomics, Expressing

Journal: Research

Article Title: Fut2 Deficiency Promotes Intestinal Stem Cell Aging by Damaging Mitochondrial Functions via Down-Regulating α1,2-Fucosylation of Asah2 and Npc1

doi: 10.34133/research.0343

Figure Lengend Snippet: Loss of α1,2-fucosylation of Asah2 and Npc1 induced stemness impairment and mitochondrial dysfunction in ISCs. (A) The protein level of Npc1, Asah2, and Bsg in whole-cell lysate and UEA-I-enriched proteins of WT and corresponding SDM organoids. (B) Images of WT, Asah2-N444Q, and Npc1-N961Q organoids and IF analysis of EGFP-labeled ISCs. Scale bars, 100 μm. (C) EdU assays in WT, Asah2-N444Q, and Npc1-N961Q organoids. Scale bar, 100 μm. (D) qPCR results of stemness markers in WT, Asah2-N444Q, and Npc1-N961Q organoids. (E to G) SA-β-gal staining of senescent cells, MitoSOX-indicated mtROS, and JC-1-indicated MMP analysis in WT, Asah2-N444Q, and Npc1-N961Q organoids. Scale bars, 100 μm. (H) Activity assays of respiratory complexes I, III, IV, and V in WT and Asah2-N444Q organoids. (I) Protein expression of mTOR and p-mTOR in WT and Npc1-N961Q organoids. (J) Expression of mitochondrial and mitophagy proteins in the whole-cell lysate of WT and Npc1-N961Q organoids. CCCP was used to activate mitophagy. (K) Expression of mitochondrial and mitophagy proteins in lysosome fractions of WT and Npc1-N961Q organoids. * P < 0.05 and ** P < 0.01.

Article Snippet: Ten micrograms of Asah2 recombinant protein (MedChemExpress, HY-P76735) and 250 nM mTOR inhibitor Torin1 (MedChemExpress, HY-13003) were used to treat organoids to confirm the role of Asah2 and Npc1 fucosylation.

Techniques: Labeling, Staining, Activity Assay, Expressing

Journal: Research

Article Title: Fut2 Deficiency Promotes Intestinal Stem Cell Aging by Damaging Mitochondrial Functions via Down-Regulating α1,2-Fucosylation of Asah2 and Npc1

doi: 10.34133/research.0343

Figure Lengend Snippet: Asah2 supplement and mTOR inhibition ameliorated stemness impairment and mitochondrial dysfunction in Fut2 ΔISC organoids. (A) Development of organoids derived from WT and Fut2 ΔISC mice, with or without Asah2 and Torin1 treatment. Scale bar, 100 μm. (B) Statistical analysis of surface areas and budding number of organoids. (C and D) IF analysis of EGFP and EdU in WT and Fut2 ΔISC organoids, with or without Asah2 and Torin1 treatment. Scale bars, 100 μm. (E and F) JC-1-indicated MMP and MitoSOX-indicated mtROS analysis in WT and Fut2 ΔISC organoids, with or without Asah2 and Torin1 treatment. Scale bars, 100 μm. (G) Activity of complexes I, III. IV, and V in WT, Fut2 ΔISC, and Fut2 ΔISC + Asah2 organoids. (H) Expression of mitochondrial and mitophagy proteins in whole-cell lysate of Fut2 ΔISC and Fut2 ΔISC + Torin1 organoids. CCCP was used to activate mitophagy. (I) Expression of mitochondrial and mitophagy proteins in lysosome fractions of Fut2 ΔISC and Fut2 ΔISC + Torin1 organoids. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Ten micrograms of Asah2 recombinant protein (MedChemExpress, HY-P76735) and 250 nM mTOR inhibitor Torin1 (MedChemExpress, HY-13003) were used to treat organoids to confirm the role of Asah2 and Npc1 fucosylation.

Techniques: Inhibition, Derivative Assay, Activity Assay, Expressing

Journal: Research

Article Title: Fut2 Deficiency Promotes Intestinal Stem Cell Aging by Damaging Mitochondrial Functions via Down-Regulating α1,2-Fucosylation of Asah2 and Npc1

doi: 10.34133/research.0343

Figure Lengend Snippet: The mechanism of Fut2-deficiency-induced facilitation of ISC aging. Fut2 knockout resulted in loss of α1,2-fucosylation on Asah2 and Npc1, which impaired respiratory complexes and mitophagy, and therefore suppressed ATP production and promoted ROS production. These mitochondrial dysfunctions promoted stemness impairment and ISC aging. This figure is illustrated using BioRender ( www.biorender.com ).

Article Snippet: Ten micrograms of Asah2 recombinant protein (MedChemExpress, HY-P76735) and 250 nM mTOR inhibitor Torin1 (MedChemExpress, HY-13003) were used to treat organoids to confirm the role of Asah2 and Npc1 fucosylation.

Techniques: Knock-Out