Journal: Stem Cell Reviews and Reports

Article Title: The Treasury of Wharton's Jelly

doi: 10.1007/s12015-021-10217-8

Figure Lengend Snippet: Flow cytometry. A typical example of FACS based expression analysis for transcription factors (upper histograms) and surface receptors (lower histograms) of VSELs. Each histogram compares given antibodies (given in black numbers) and specifc isotype controls (given in red numbers). Percentage of positive VSELs and the median fluorescence were assessed

Article Snippet: For fixation, permeabilization and staining procedures buffers by “Transcription Factor Staining Buffer Set” by Miltenyi Biotec (Berglisch Gladbach, Germany) were used.

Techniques: Flow Cytometry, Expressing, Fluorescence

Journal: Stem Cell Reviews and Reports

Article Title: The Treasury of Wharton's Jelly

doi: 10.1007/s12015-021-10217-8

Figure Lengend Snippet: Expression of pluripotency associated marker proteins examined by Immunofluorescence staining. VSELs were separated from UC-MSCs mass population, the tiny cells are between 5 and 7 μm and surrounded with dark spots ( a ). VSEL suspension cells were stained extracellular with anti-bodies against SSEA-4, CD184, and CD133 followed by nuclear staining of transcription factors Nanog, Oct-4, and Sox-2. Expression of each marker was detected. The nuclei were stained with HOECHST. Weak background Fluorescence signals of isotype controls were subtracted. The scale bars represented 50 μm ( b ). The magnification for SSEA-4/Sox-2 and CD133/Oct-4 was 20-fold and for CD184/Nanog 10-fold

Article Snippet: For fixation, permeabilization and staining procedures buffers by “Transcription Factor Staining Buffer Set” by Miltenyi Biotec (Berglisch Gladbach, Germany) were used.

Techniques: Expressing, Marker, Immunofluorescence, Staining, Suspension, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: SC transplantation shifted the CD11b immune cell population from an Arg1 − iNOS + pro-inflammatory to an intermediate Arg1 + iNOS + phenotype after SCI. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) show, compared with SCI controls ( A – C ), a decreased percentage of CD11b cells stained with Arg1 − iNOS + and an increased percentage for Arg1 + iNOS + in animals receiving SC transplants ( D – F ). Results are expressed as mean ± standard deviation (SD). Abbreviations on the graphs are: Fluorescein isothiocyanate (FITC), Allophycocyanin (APC) and Forward Scatter (FSC-A). For panels ( B , E ), the blue dots represent the CD11b population that is iNOS − -Arg1 − , the orange dots represent the CD11b population that is iNOS + -Arg1 − and the green dots represent the CD11b population that is double positive for both iNOS + -Arg1 + . These colored dots are also shown in the forward scatter plots of panels ( A , D ).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Transplantation Assay, Flow Cytometry, Staining, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: CD68 immune cells with a pro-inflammatory Arg1 − iNOS + phenotype after SCI are reduced by SC transplantation. Representative images of flow cytometry analysis and pie charts of CD68 population dynamics at 14 days post-injury (7 days post-transplantation) reveal that CD68 immune cells with a pro-inflammatory Arg1 − iNOS + phenotype after SCI ( A – C ) are reduced by the intraspinal transplantation of SCs ( D – F ). Results are expressed as mean ± SD. For panels ( A , D ), the orange dots represent the ED1 population that were gated based on their forward and side scatter from the total events (blue) that were acquired. The orange dots in the different quadrants of ( B , E ) represent the ED1 population that was positive for either Arg1 (top left quadrant) or iNOS (bottom right quadrant) or double positive for both markers (top right quadrant).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Transplantation Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: CD11b immune cells expressing a highly pro-inflammatory CD38 + iNOS + phenotype or anti-inflammatory Arg1 + CD163 + form after SCI were unaltered by SC transplantation. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) reveal that CD11b immune cells with a highly pro-inflammatory CD38 + iNOS + phenotype after SCI ( A , B ) are not altered by SC transplantation ( E , F ). Similarly, CD11b immune cells with a highly anti-inflammatory Arg1 + CD163 + phenotype were unchanged across SCI controls ( C , D ) and SC-transplanted groups ( G , H ). Results are expressed as mean ± SD. For panels ( A , B ), the blue dots represent the CD11b population that were CD38 − iNOS − , the orange dots represent the CD11b population that were CD38 − iNOS + , the gray dots represent the CD11b population that were CD38 + iNOS − and the green dots represent the CD11b population that were double positive for both CD38 + iNOS + . For panels ( C , D ), the configuration for the colored dots is the same for the representation labeling, single, double or absent, though the proteins Arg1 and CD163 are represented rather than CD38 and iNOS.

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Expressing, Transplantation Assay, Flow Cytometry, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: Cytokine profile of CD11b + cells is unaltered by SC implants after SCI. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) show that CD11b immune cells have unaltered expression of pro- (TNF-α, IL-1β) and anti-inflammatory (IL-4, IL-10) cytokines comparatively among SCI controls ( A – E ) and SC-transplanted groups ( F – J ). Results are expressed as mean ± SD. For panels ( A – D , F – I ) the blue dots represent the cell population that were CD11b − or did not express the selected cytokines, whereas the green dots represent the CD11b + population that expressed CD11b and the cytokine of interest (top right quadrant).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Flow Cytometry, Transplantation Assay, Expressing