Journal: PLoS ONE

Article Title: Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain

doi: 10.1371/journal.pone.0186717

Figure Lengend Snippet: (A)-(C): fluorescent microscopy of a hMSC culture 24 hours after labeling with MC03F microparticles. (A) MC03F microparticles (Dragon Green fluorescence). (B) Cell nuclei (DAPI blue fluorescence). (C) A and B merged. (D) Transmitted light microscopy of unstained hMSC culture (the same area as in A-C) 24 hours after labeling with MC03F microparticles. SPIO microparticles are visualized in the cytoplasm as brown spots around clear nuclei. (E) Flow cytometry analysis of cells 24 hours after labeling. The solid line presents data for labeled hMSCs and the dotted line—for unlabeled, control hMSCs. X-axis shows fluorescence intensity and Y-axis—cell counts. The plot demonstrates that about 96% of the cells contained Dragon Green fluorescent microparticles. (F) Influence of the labeling with MC03F microparticles on hMSCs viability and proliferation. Optical density (Y axis) is proportional to lactate dehydrogenase (LDH) activity in the cells and, hence, to the number of living cells. The presented histograms show that the numbers of living cells were not significantly different in labeled and control cultures indicating the absence of negative effects associated with labeling on cell viability and proliferation. The scale bars on all microphotographs mark 100 μm.

Article Snippet: Byun et al.[ ] , hMSCs from bone marrow—Feridex® SPIO nanoparticles (Bayer Healthcare Pharmaceuticals, USA) , Clinical 3T - SWI , 6.3x10 4 / -.

Techniques: Microscopy, Labeling, Fluorescence, Light Microscopy, Flow Cytometry, Control, Activity Assay

Journal: PLoS ONE

Article Title: Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain

doi: 10.1371/journal.pone.0186717

Figure Lengend Snippet: (A) The medians of the values of the ratio of minimum to mean signal calculated from data obtained using different pulse sequences after injection of 10 1 , 10 2 or 10 3 of SPIO-labeled hMSCs or saline into the rat striatum. Data for cell concentrations higher than 10 3 are not shown on the graph, because for them the minimum signal was zero for most MRI pulse sequences. Whiskers on the plot represent interquartile range. (B) Volumes of the hypointensity zones calculated from SWI data obtained after injection of varying quantities of SPIO-labeled hMSCs or saline into the rat striatum (box and whisker plot). (C) Values of the ratio of minimum to mean signal calculated from SWI data obtained after injection of 10 1 , 10 2 or 10 3 of SPIO-labeled hMSCs or saline into the rat striatum (box and whisker plot). Greek letters show statistical significance of differences between pulse sequences (A), or saline-injected and SPIO-labeled cell-transplanted rats (B, C): α –p<0.05; β –p<0.01; γ –p<0.001.

Article Snippet: Byun et al.[ ] , hMSCs from bone marrow—Feridex® SPIO nanoparticles (Bayer Healthcare Pharmaceuticals, USA) , Clinical 3T - SWI , 6.3x10 4 / -.

Techniques: Injection, Labeling, Saline, Whisker Assay

Journal: PLoS ONE

Article Title: Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain

doi: 10.1371/journal.pone.0186717

Figure Lengend Snippet: (A) MRI of rat brain (T2WI and SWI) immediately after intracerebral injection of 10 5 hMSCs double-labeled with MC03F SPIO microparticles and PKH26. Hypointense regions indicate the location of SPIO labeled cells or probably extracellular SPIO microspheres. (B) High-magnification confocal micrographs of the hMSCs injection site. The same rat brain as in fig A. White arrowhead points to a double-labeled hMSC (membrane dye PKH26 is red, SPIO microparticles in the cytoplasm are green, and cell nucleus stained with DAPI is blue). Single clusters of extracellular iron can also be visualized (white arrows). The scale bars represent 10 μm. (C) Confocal panoramic micrographs of the hMSCs injection site (the same rat brain as in fig A). Labeled hMSCs are located along the track of the injection needle and in corpus callosum. The scale bars represent 500 μm. (D) MRI of rat brain (T2WI and SWI) after intra-arterial transplantation of 10 5 hMSCs labeled with SPIO-microparticles and PKH26. The white arrows on the SWI picture indicate the location of SPIO labeled cells. (E) High-magnification confocal micrographs (the same rat brain as in fig D) of transplanted hMSCs. White arrowhead points at double-labeled cells (membrane dye PKH26 is red, SPIO microparticles in the cytoplasm are green, and cell nuclei stained with DAPI is blue). The scale bars represent 20 μm.

Article Snippet: Byun et al.[ ] , hMSCs from bone marrow—Feridex® SPIO nanoparticles (Bayer Healthcare Pharmaceuticals, USA) , Clinical 3T - SWI , 6.3x10 4 / -.

Techniques: Injection, Labeling, Membrane, Staining, Transplantation Assay

Journal: PLoS ONE

Article Title: Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain

doi: 10.1371/journal.pone.0186717

Figure Lengend Snippet: MRI detection thresholds of SPIO-labeled stem cells in phantom studies.

Article Snippet: Byun et al.[ ] , hMSCs from bone marrow—Feridex® SPIO nanoparticles (Bayer Healthcare Pharmaceuticals, USA) , Clinical 3T - SWI , 6.3x10 4 / -.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain

doi: 10.1371/journal.pone.0186717

Figure Lengend Snippet: MR detection thresholds of SPIO-labeled stem cells in live animal brain.

Article Snippet: Byun et al.[ ] , hMSCs from bone marrow—Feridex® SPIO nanoparticles (Bayer Healthcare Pharmaceuticals, USA) , Clinical 3T - SWI , 6.3x10 4 / -.

Techniques: Sequencing

Journal: Biomarkers in Cancer

Article Title: Differential Expression of Cysteine Dioxygenase 1 in Complex Karyotype Liposarcomas

doi: 10.4137/BIC.S14683

Figure Lengend Snippet: CDO1 expression during adipogenic differentiation of hMSCs. ( A ) TG content increases during in vitro differentiation of hMSCs relative to LS2, a PLS cell line. ( B ) Quantitative RT-PCR analysis of CDO1 transcript level during adipogenic differentiation of hMSCs. CDO1 expression remains low until terminal differentiation. Values are relative to CDO1 expression in mature adipocytes. ( C ) Indirect immunofluorescence for CDO1 (shown in red). Nuclei were counterstained with DAPI (blue). CDO1 protein was increased at the later stages of induction, particularly following the fourth induction cycle (scale bar = 100 μm).

Article Snippet: Adipogenic differentiation was induced in Poietics bone marrow-derived hMSCs (Lonza) according to the manufacturer’s instructions.

Techniques: Expressing, In Vitro, Quantitative RT-PCR, Immunofluorescence

Journal: Tissue Engineering. Part A

Article Title: Pushing the Envelope in Tissue Engineering: Ex Vivo Production of Thick Vascularized Cardiac Extracellular Matrix Constructs

doi: 10.1089/ten.tea.2014.0477

Figure Lengend Snippet: Compartmentalized dynamic recellularization using monocultures of human mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs). A functioning perfusion chamber can be trans-located from the CO 2 incubator into a biological cabinet where sterile handling is available (a) . Using this system, decellularized thick pcECM scaffolds (b) regain full thickness appearance after 48 h of perfusion, as viewed from top or side (c, d, respectively ) . H&E staining 7 days postseeding (e) . Cell survival when cultivated under various physiological flow rates, using different seeding times (1.5 or 24 h), determined after 24 h of perfusion (f) . * Denotes significantly different results p <0.05. Transferring of statically cultivated thick constructs ( t =30 days, marked with an arrow ) to further cultivation in the dynamic system exhibits a significant ( p <0.05) increase in cell quantities (g) . Dashed red line represents the 95% confidence interval of the mean. H&E staining of histological cross-sections, 7 days postdynamic cultivation of hMSCs seeded through the bulk of the pcECM by injection—fibers are shown in red and cell nuclei in blue (h) . Specific antibody staining for CD44 suggests that the hMSCs are anchored to the pcECM through their HA receptors (i) . Live confocal imaging (hMSCs stained with Hoechst) of the endocardial surface after 21 days of static culture reveals densely populated surfaces in accordance with the mathematical model prediction of steady state densities (j) . Re-endothelialization of the vascular network within the pcECM is demonstrated using a monoculture of HUVEC-GFPs ( green ) forming 14 days postseeding and perfusion, a monolayer coating in a cobble stone-like formation (k) . Cross-section staining of the GFP expressing cells ( green ) with CD31 ( red ) demonstrated endothelium formation within the lumen of the blood vessel (l) . In all experiments, results represent three biological repetitions ( n =3). Scale bars: (e) , (h) , (j–l) , 100 μm; (i) , 50 μm. Color images available online at www.liebertpub.com/tea

Article Snippet: Bone marrow hMSCs were purchased from Lonza and cultured in humidified incubator at 37°C and 5% CO 2 using alpha modified Eagle's medium (α-MEM; Biological Industries) supplemented with 20% fetal bovine serum, 1% Pen-Strep, and 0.4% Fungizone ® .

Techniques: Sterility, Staining, Transferring, Construct, Injection, Imaging, Expressing

Journal: Nature Communications

Article Title: Mechanically-sensitive miRNAs bias human mesenchymal stem cell fate via mTOR signalling

doi: 10.1038/s41467-017-02486-0

Figure Lengend Snippet: Modulation of miRNA activity can modulate the hMSC differentiation bias in response to substrate stiffness. Differentiation bias of hMSCs treated with a miRNA mimics or b miRNA inhibitors and cultured on 70 kPa gels and c treated with miRNA mimics or d miRNA inhibitors and cultured on 0.6 kPa gels. Data is shown as mean ± SD of the change in proportion of osteoblasts (see Supplementary Fig. for additional donors). Samples were analysed by one-way ANOVA with Tukey post hoc testing. Statistically different samples are denoted by (*)

Article Snippet: hMSCs (bone marrow-derived, Lonza) were cultured in DMEM-low glucose supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (DMEM/ps) and 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Cell Culture