Journal: Frontiers in Cell and Developmental Biology

Article Title: Senescent Tissue-Resident Mesenchymal Stromal Cells Are an Internal Source of Inflammation in Human Osteoarthritic Cartilage

doi: 10.3389/fcell.2021.725071

Figure Lengend Snippet: Serial cell culture passage of OAC induces dedifferentiation into NCSC-like cells and subsequent senescence into pro-inflammatory OA-MSC. (A) FACS analysis of CD166 positive cells during OA chondrocyte passaging in OA-MSC culture medium or OAC medium. The percentage of CD166 positive cells in each passage ( x -axis) was indicated in red. The increase of the number of large sized cells ( y -axis), a characteristic of senescent cells, was indicated by brackets in Passage 5. The data is representative of three experiments. (B) Cell morphology change of OA chondrocytes during cell culture passages in OAC medium and OA-MSC culture medium. OAC changed its morphology from cuboidal shape typical of chondrocytes (aquamarine arrows) in early passages (2 and 3) to spindle shape typical of fibroblasts (gray arrows) in mid passage (4) to large cells with processes typical of senescent cells (yellow arrows) in late passage (5). Bar = 200 μm. (C) Expression of aging related genes during OAC culture passages in OAC medium and OA-MSC medium. mRNA levels of TNFR2, CXCL1, CXCL8, P16 were quantified by real-time RT-PCR. The value of each sample group = mean ± SD. N = 3 for all groups. * p -value < 0.05; *** p -value < 0.001. (D) The components of OAC medium and OA-MSC medium.

Article Snippet: Pre-conjugated antibodies CD166-FITC and mouse-IgG-FITC were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Techniques: Cell Culture, Passaging, Expressing, Quantitative RT-PCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: Senescent Tissue-Resident Mesenchymal Stromal Cells Are an Internal Source of Inflammation in Human Osteoarthritic Cartilage

doi: 10.3389/fcell.2021.725071

Figure Lengend Snippet: Identification of OAC, NCSC-like cells, and OA-MSC in human OA articular cartilage. (A) Differential expression analysis of cytokine, chemokine, and related genes in OAC, NCSC, and OA-MSC. RNAseq analysis was performed with OAC, NCSC3, and OA-MSC2 samples ( n = 3). Yellow color represents higher copy number up to 10,000, while blue color represents lower copy number down to 0. Gray color represents intermediate copy number. Note that while IL-1β was synthesized by OA-MSC, neither OAC nor NCSC synthesized IL-1β. (B) Differential expression analysis of cytokine and chemokine receptors and related genes in OAC, NCSC, and OA-MSC. RNAseq analysis was performed with OAC, NCSC3, and OA-MSC2 samples ( n = 3). Yellow color represents higher copy number up to 30,000, while blue color represents lower copy number down to 0. Gray color represents intermediate copy number. (C) Immunofluorescence analysis of different types of cells in human OA cartilage. While chondrocytes are CD166 – , mesenchymal stromal cells are CD166 + . OAC does not synthesize IL-1β but binds to IL-1β because it expresses IL-1R. OAC is CD166 – /IL-1β – ; OAC binding to IL-1 β (OAC with cytokine) is CD166 – /IL-1β + , NCSC-like cell is CD166 + /IL-1β – , and OA-MSC is CD166 + /IL-1β + cells in OA cartilage. CD166 was labeled by rhodamine red conjugated secondary antibody. IL-1β was labeled by green fluorescein conjugated secondary antibody. The nucleus was stained with blue Hoechst dye. (D) The double immunofluorescence histochemical analysis of CD166 and IL-1β in different zones in human OA articular cartilage. CD166, a marker of MSC, was indicated by rhodamine-conjugated secondary antibody (red). IL-1β, a marker of SASP was indicated by fluorescein-conjugated secondary antibody (green). The nucleus was stained with DAPI (blue). (E) The percentages of OAC, OAC (with cyto), NCSC-like cells, and OA-MSC in different zones of human OA articular cartilage ( n = 3). Statistical analysis was presented in .

Article Snippet: Pre-conjugated antibodies CD166-FITC and mouse-IgG-FITC were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Techniques: Quantitative Proteomics, Synthesized, Immunofluorescence, Binding Assay, Labeling, Staining, Marker

Journal: Journal of Neuroinflammation

Article Title: Extended interval dosing of ocrelizumab modifies the repopulation of B cells without altering the clinical efficacy in multiple sclerosis

doi: 10.1186/s12974-023-02900-z

Figure Lengend Snippet: Antibodies for CyTOF

Article Snippet: 164Dy , CD166 , 3A6 , Standard BioTools , 0.5 , Surface.

Techniques: Concentration Assay