Journal: Scientific Reports

Article Title: Encapsulation of human limbus-derived stromal/mesenchymal stem cells for biological preservation and transportation in extreme Indian conditions for clinical use

doi: 10.1038/s41598-019-53315-x

Figure Lengend Snippet: Phenotypic expression of the biomarkers. Immunostaining of the encapsulated hLSMCs under transit for 3 days: Alginate encapsulated hLSMCs of both groups, stored/under transit for 3 days have shown the expression of Pax6 + , stem-cell biomarkers (ABCG2 + , p63-α + ) and the mesenchymal biomarkers (VIM + , CD90 + , CD105 + and CD45 − ) with respect to the control cells. Blue: DAPI, nuclear stain. Scale: 100 µM.

Article Snippet: The antibody panel was composed of (a) ABCG2 (1:100, 18841, Santa Cruz Biotechnology, USA), Pax6 (1:300, 901301, BioLegend, USA), p63-α (1:100, 4892S, Cell Signalling technology, USA) and Col-III (1:100, ab7778, Abcam, UK), as positive markers of the human limbal stem cell phenotype; HLA-DR (1:100, ab55152, Abcam, UK), and CD45 (1:100, 13197, Cell Signalling Technology, USA) as negative marker for mesenchymal origin, (b) CD73 (1:100, 13160, Cell Signalling Technology, USA), CD105 (1:100, 376381, Santa Cruz Biotechnology, USA), and VIM (1:100, 6260, Santa Cruz Biotechnology, USA) as positive markers of the mesenchymal phenotype.

Techniques: Expressing, Immunostaining, Control, Staining

Journal: Scientific Reports

Article Title: Encapsulation of human limbus-derived stromal/mesenchymal stem cells for biological preservation and transportation in extreme Indian conditions for clinical use

doi: 10.1038/s41598-019-53315-x

Figure Lengend Snippet: Quantification of the gene expression using real-time PCR. Immunostaining of the encapsulated hLSMCs under transit for 5 days: Alginate encapsulated hLSMCs stored at 4 °C did not show expression of the stem-cell (ABCG2 − ). The RT group cells have showed similar phenotype as the control group (ABCG2 + , Pax6 + p63-α + , VIM + , CD90 + , CD105 + , CD45 − , HLADR + , Col-III + , and CD73 + ). Blue: DAPI, nuclear stain. Scale : 100 µM.

Article Snippet: The antibody panel was composed of (a) ABCG2 (1:100, 18841, Santa Cruz Biotechnology, USA), Pax6 (1:300, 901301, BioLegend, USA), p63-α (1:100, 4892S, Cell Signalling technology, USA) and Col-III (1:100, ab7778, Abcam, UK), as positive markers of the human limbal stem cell phenotype; HLA-DR (1:100, ab55152, Abcam, UK), and CD45 (1:100, 13197, Cell Signalling Technology, USA) as negative marker for mesenchymal origin, (b) CD73 (1:100, 13160, Cell Signalling Technology, USA), CD105 (1:100, 376381, Santa Cruz Biotechnology, USA), and VIM (1:100, 6260, Santa Cruz Biotechnology, USA) as positive markers of the mesenchymal phenotype.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Immunostaining, Expressing, Control, Staining

Journal: Scientific Reports

Article Title: Encapsulation of human limbus-derived stromal/mesenchymal stem cells for biological preservation and transportation in extreme Indian conditions for clinical use

doi: 10.1038/s41598-019-53315-x

Figure Lengend Snippet: Tabular format denoting the number of cells showing positive expression of the phenotypic biomarkers.

Article Snippet: The antibody panel was composed of (a) ABCG2 (1:100, 18841, Santa Cruz Biotechnology, USA), Pax6 (1:300, 901301, BioLegend, USA), p63-α (1:100, 4892S, Cell Signalling technology, USA) and Col-III (1:100, ab7778, Abcam, UK), as positive markers of the human limbal stem cell phenotype; HLA-DR (1:100, ab55152, Abcam, UK), and CD45 (1:100, 13197, Cell Signalling Technology, USA) as negative marker for mesenchymal origin, (b) CD73 (1:100, 13160, Cell Signalling Technology, USA), CD105 (1:100, 376381, Santa Cruz Biotechnology, USA), and VIM (1:100, 6260, Santa Cruz Biotechnology, USA) as positive markers of the mesenchymal phenotype.

Techniques: Expressing, Control, Biomarker Discovery

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Restore Endothelial Integrity and Alleviate Emotional Impairments in a Diabetic Mouse Model via Inhibition of MMP-9 Activity

doi: 10.3390/ijms26073355

Figure Lengend Snippet: Figure 2. DM mice exhibit synaptic deficits and increased neuroinflammation in the HIP. (A) Repre- sentative WB images showing the expression levels of the presynaptic protein synapsin I (SYN1) and the postsynaptic protein postsynaptic density protein 95 (PSD95) in the HIP of CTL and DM mice. β-actin was used as an internal control. (B) Semi-quantitative analysis of PSD95 and SYN1 expression levels from immunoblot experiments. n = 6. (C) Representative images of Iba1 immunostaining in the HIP of CTL and DM mice. The area within the dashed box is magnified and displayed in the insets. Scale bars: 100 µm (main images) and 20 µm (insets). (D) Quantitative analysis of Iba1-positive microglia in the HIP of CTL and DM mice. n = 5. (E) qPCR analysis of iNOS and Arg1 mRNA levels in the HIP of CTL and DM mice. n = 6. (F) qPCR analysis of IL-6, IL-1β, and TNF-α mRNA levels in the HIP of CTL and DM mice. n = 6. (G) ELISA measuring IL-6, IL-1β, and TNF-α levels in the serum of CTL and DM mice. n = 3. Data are presented as mean ± SEM. Statistical significance was determined using a one-tailed unpaired t-test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were blocked and incubated overnight with the following antibodies: PSD95 (SYSN, Toronto, ON, Canada, 124002, 1:1000); SYN1 (SYSN, 106011, 1:1000); Cldn5 (Invitrogen, 35-2500, 1:1000); Ocln (Invitrogen, 71-1500, 1:1000); and MMP9 (Boster, Shanghai, China; PB9669, 1:1000).

Techniques: Expressing, Control, Western Blot, Immunostaining, Enzyme-linked Immunosorbent Assay, One-tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Restore Endothelial Integrity and Alleviate Emotional Impairments in a Diabetic Mouse Model via Inhibition of MMP-9 Activity

doi: 10.3390/ijms26073355

Figure Lengend Snippet: Figure 6. MSC treatment attenuated DM-induced neuroinflammation and synaptic deficits. (A) Representative immunofluorescence images of Iba1 staining in the HIP of mice from the CTL, MSC, DM, and DM+MSC groups. The dashed box indicates the magnified region shown in the insets. Scale bars: 100 µm (main images) and 20 µm (insets). (B) Quantitative analysis of Iba1+ microglia in the HIP. n = 4. (C) qPCR analysis of iNOS and Arg1 mRNA levels in the HIP of mice from the CTL, MSC, DM, and DM+MSC groups. n = 9. (D) qPCR analysis of mRNA levels of IL-6, IL-1β, and TNF-α in the HIP of mice from the CTL, MSC, DM, and DM+MSC groups. n = 9. (E) ELISA results showing serum levels of IL-6, IL-1β, and TNF-α in CTL, MSC, DM, and DM+MSC mice. n = 4. (F) Representative WB images of PSD95 and SYN1 expression in the HIP. β-actin was used as a loading control. (G) Semi-quantitative analysis of PSD95 and SYN1 protein levels. n = 6. Data are presented as mean ± SEM. Two-way ANOVA followed by Tukey’s post hoc test. ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were blocked and incubated overnight with the following antibodies: PSD95 (SYSN, Toronto, ON, Canada, 124002, 1:1000); SYN1 (SYSN, 106011, 1:1000); Cldn5 (Invitrogen, 35-2500, 1:1000); Ocln (Invitrogen, 71-1500, 1:1000); and MMP9 (Boster, Shanghai, China; PB9669, 1:1000).

Techniques: Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Control