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: International Journal of Molecular Sciences

Article Title: High Adenosine Extracellular Levels Induce Glioblastoma Aggressive Traits Modulating the Mesenchymal Stromal Cell Secretome

doi: 10.3390/ijms21207706

Figure Lengend Snippet: Effects of ADO in the BM-MSC and GBM cell cross-talk. ( A ) The image depicts the contact-independent transwell cultures with BM-MSCs in the upper chamber and U343MG in the lower chamber. U343MG cell proliferation was evaluated after 24 and 48 h with Crystal violet, as reported in . ( B ) The image depicts the contact-independent transwell cultures with U343MG in the upper chamber and BM-MSCs in the lower chamber. BM-MSCs cell proliferation was evaluated after 48 h with Crystal violet, as reported in . ( C ) U343MG cells were seeded in the upper chamber and BM-MSCs treated with ADO (100 nM and 100 µM) in the lower chamber. After 24 h, U343MG cell invasion was analyzed using Matrigel basement membrane transwell system, as described in . Representative images are shown. Cell migration was quantified by counting the number of cells that migrated into the lower surface of the transwell membrane. The data are reported as percentage of cell invasion versus the CTRL set to 100% and are the means ± SEM of three independent experiments. The significance of the differences was determined by one-way ANOVA, followed by Bonferroni’s post-hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTRL.

Article Snippet: Human bone marrow MSCs (BM-MSCs) and glioblastoma stem cells (U343MG) were purchased by CLS (CLS Cell Lines Service GmbH, Eppelheim, Germany).

Techniques: Migration

Journal: PLOS ONE

Article Title: Recovery after human bone marrow mesenchymal stem cells (hBM-MSCs)-derived extracellular vesicles (EVs) treatment in post-MCAO rats requires repeated handling

doi: 10.1371/journal.pone.0312298

Figure Lengend Snippet: (a–c) Characterization and quantification of human-bone marrow mesenchymal stem cell (hBM-MSC)-derived extracellular vesicles (EVs). (a) Nanosight Tracking analyses (NTA) showing the size distribution pattern of EVs. (b) Western blots of hBM-MSCs marker (GM130) and EVs markers (ALIX, CD9, CD81). “Wash” lane represents the negative control. Each image comes from different blots. (c) Representative image of EVs using transmission electron microscopy (TEM). Scale bar 100 nm. (d–f) In vivo tracking of ExoGlow-labeled EVs demonstrated that EVs reach the brain and accumulate near the stroke injury 6 hours after administration at 2 dps. (d) Timeline of intranasal single dose treatment of ExoGlow-labeled EVs (~2.4 x 10 9 EVs in 200 μl), starting 2 dps and collecting the tissue 6 hours after. (e) Representative coronal section after a single intranasal administration of ExoGlow-labeled EVs (~2.4 x 10 9 EVs in 200 μl) at 2 dps. Hoechst indicates cell nuclei. Scale bars 4 mm and 100 μm. (f) Quantification of the average size of ExoGlow+ particles in the ipsilateral and contralateral hemispheres. Data was analyzed using a paired t-test (*p<0.05). Bars show mean +/− SD. Each symbol represents the average size of ExoGlow+ particles from all ROIs in each hemisphere (5 ROIs/hemisphere in each rat) of a single rat ( n = 4 ).

Article Snippet: Human bone marrow mesenchymal stem cells (hBM-MSCs) were purchased from Celprogren (frozen vial with ~1.2 x 10 6 cells; #36094–22) and plated on human mesenchymal bone marrow stem cell culture extracellular expansion matrix pre-coated T75 Flasks (Celprogen, #E36094-21-T75) following Celprogren recommendations.

Techniques: Derivative Assay, Western Blot, Marker, Negative Control, Transmission Assay, Electron Microscopy, In Vivo, Labeling

Journal: Nature Communications

Article Title: DNA-templated synthesis of biomimetic cell wall for nanoencapsulation and protection of mammalian cells

doi: 10.1038/s41467-019-10231-y

Figure Lengend Snippet: Evaluation of shielding enhancement. a Examination of cell viability at different time points. The viability of native cells was used as 100%. 0 h: analysis of the cells right after the synthesis of the template or BCW. b Schematic comparison of the cells covered with (bottom) or without (top) BCW when exposed to environmental assaults. c Shielding enhancement-centrifugal force relationship. After the assault, the cells were cultured and examined with the LIVE/DEAD staining. Green: live; red: dead. Representative images were taken after the cells were exposed to the centrifugal force of 6200× g . d Shielding enhancement-osmotic imbalance relationship. The cells were stained with Calcein-AM (green) for imaging before exposed to the conditions of osmotic imbalance. Representative images were taken after the cells were incubated in the solution of 0.4% NaCl. CCRF-CEM was used for Fig. 5a–d. e Shielding enhancement-immune attack relationship. K562 and NK-92MI were used as target and effector cells, respectively. Representative images were taken at the 2.5:1 effector/target ratio. Green: CFSE. Red: dead cells. Scale bars: 50 μm. Data are presented as mean ± standard deviation as indicated by error bars ( n = 3). f In vivo imaging of human bone marrow MSCs subcutaneously transplanted into the mice. MSCs expressed red fluorescence protein (RFP). BF, bright field. (−): naked cells (without BCW); (+): BCW-covered cells. Color-coding of fluorescence intensity in arbitrary units (a.u.) is equally scaled for the images. g Staining of endothelial cells using a mouse-specific anti-CD31 antibody. ** p < 0.01. The paired Student’s t -test was used to compare the two groups. Scale bars: 200 μm. Data are presented as mean ± standard deviation as indicated by error bars ( n = 6)

Article Snippet: RFP-Tagged human bone marrow derived MSCs (RFP-MSCs) were purchased from Angio-Proteomie (Boston, MA) and maintained in stem cell growth medium (Lonza).

Techniques: Comparison, Cell Culture, Staining, Imaging, Incubation, Standard Deviation, In Vivo Imaging, Fluorescence