Cd90 fitc

FLSs were isolated and cultured from RA synovium. Tissue biopsies were finely minced into pieces and transferred to a tissue culture flask in Dulbecco's modified Eagle's medium (DMEM) (Gibco Laboratories, Invitrogen, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco Laboratories). Within 14 days, FLSs migrated out from the tissue explant and were grown to approximately 95% confluency. FLSs were subsequently trypsinized, collected, re-suspended, and planted for proliferation. FLSs from passages 3–5 were used for the following experiments.
The morphological characters of FLSs were observed under the light microscope. The expression level of surface markers on FLSs were detected for characterization using flow cytometry. FLSs from passages 3 were trypsinized, centrifuged, and stained with commercial monoclonal antibodies CD68FITC, CD14FITC, CD90FITC, CD55PE (Biolegend, USA) for 20 min. And isotype-matched control antibodies were used as methodologic controls. Stained cells were subsequently analyzed using a FACSCalibur Flow Cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). For each analysis, 10,000 events were evaluated with the software FlowJo 7.6 (Becton Dickinson, Franklin Lakes, NJ, USA).

Cells were detached with Accutase (Sigma-Aldrich; 5 min at 37 °C) and then incubated with the following mouse anti-human mAbs for 30 min on ice: CD73-Pacific Blue™, CD90-FITC, CD105-FITC, CD166-PE, CD34-APC, CD45-PerCP, CD117-PE (all from BioLegend) and CD133-APC (Miltenyi Biotec). After two washing steps (PBS with 2 % FBS), the sample tubes were acquired on a BD FACSAria III (BD Biosciences) and at least 10.000 events were recorded. Data were analyzed with BD FACSDiva and FlowJo V10 software.

Sal B was purchased from Maclean’s Biochemistry (China), dissolved in DMSO, and stored in the refrigerator at −80°C before use, The annexin V/FITC apoptosis kit was purchased from KGI (China), fetal bovine serum was purchased from Clark Bioscience (United States), DMEM/F12 was purchased from Biosharp (China), HAMA-400K hydrogel was purchased from Suzhou Yongqinquan Intelligent Equipment Co., Ltd. (China), CD105-APC purchased from EXBIO (Czech Republic), CD34-FITC antibody was purchased from Santa Cruz Biotechnology (United States), CD45-PE was purchased from Invitrogen (United States), CD90-FITC was purchased from Biolegend. JAK2, p-JAK2, STAT3, p-STAT3, BAX, and Bcl2 antibodies were purchased from Affinity Biosciences (China), DAPI and FITC-labeled ghost cyclic peptides were purchased from Sloarbio (China), and the Calcein-AM/PI Double Stain Kit was purchased from Yeasen Biotechnology (China), Sprague-Dawley (SD) rat bone marrow MSC osteogenesis, adipogenesis, and chondrogenesis differentiation kits were purchased from Cyagen Biotech (China).
The SD rats used in this experiment were purchased from Hunan Sleek Jingda Laboratory Animal Co., Ltd. (license number SCXK (Xiang) 2019-0004) and approved by the Ethics Committee of Bengbu Medical College (approval number: Lunde Keji [2020] No. 198).

The decidual unsorted cells (dUCs; including dCD34⁺ and dCD34^‐ cells) were collected and labelled with the following antibodies for dual staining: CD34‐FITC (BD, Franklin Lakes, NJ, USA)/ c‐kit‐PE (BD) for 30 minutes at 4°C. The dCD34⁺ cells were stained with CD34‐FITC (BD), c‐kit‐PE (BD), CD90‐FITC (BioLegend, San Diego, CA, USA), CD105‐APC (BioLegend), CD31‐FITC (BD), VEGFR‐2‐FITC (BD), VE‐cadherin‐FITC (BD), HLA‐ABC‐FITC (Abcam) and HLA‐DR‐FITC (Abcam). Then, the cells were washed three times with cold PBS before being centrifuged at 1000 rpm for 5 minutes. Immunoreactivity of the cell surface antibody markers was assayed by fluorescence‐activated cell sorting (FACS; BD).