Cd105 fitc

Wharton's Jelly-derived MSCs, treated as described above, were subjected to flow cytometric analysis at passages 1–10. Antibodies to CD44-FITC, CD45-FITC, CD73-FITC, CD90-FITC, and CD105-FITC (Abcam, USA) were used to mark cell surface epitopes. All analyses were standardised against negative control cells incubated with isotype-specific IgG1-FITC (Santa Cruz). The number of cells staining positive for a marker was determined by the percentage of cells present within a gate established using the FITC-conjugated isotype-matched control. At least 50 000 events were acquired on a FACScan cell sorter (Becton Dickinson, NJ, USA). For osteogenic and chondrogenic differentiation, MSCs were incubated with the respective media (Life Technologies) for up to 21 days. The medium was replaced twice every week. For osteogenic differentiation mineralization was visible as red stained calcium deposition. For chondrogenic differentiation, proteoglycans were stained with toluidine blue O which was visible as purple. For adipogenic differentiation, Oil Red O staining was done and lipid droplets were quantified.

PVASCs at passages 3–6 were analyzed using a fluorescence-activated cell sorter (FACS). Briefly, cells were incubated for 30 min at 4°C with the following antibodies: Sca1-Percp (abcam); CD105-FITC (abcam); CD90 (Invitrogen); CD45-FITC (abcam); and CD31-PE (BD Biosciences). The cells without any staining were served as blank. Flow cytometry was performed on a BD flow cytometer. FCS files were exported and analyzed using FlowJo V10 software.
The adipogenic differentiation medium, chondrogenic differentiation medium, and osteogenic differentiation medium were purchased from CHEM. PVASCs were seeded on gelatin-coated plates and then cultured with a specific differentiation medium and maintained for 2 weeks. The medium was changed every 3 days. After 2 weeks, cells were stained with Alcian Blue, Oil red O, or Alizarin Red S(CHEM) following the manufacturers’ instructions, respectively, to assess adipogenic, chondrogenic, and osteogenic differentiation.
PVASCs are positive for Sca1, CD90, and CD105 and negative for CD45 and CD31 (Supplementary Figures 1A,B). PVASCs were capable of differentiating into adipocytes, chondrocytes, and osteoblasts, respectively (Supplementary Figures 1C–E).

BM‐MSCs were purchased from the Cell Center of the Chinese Academy of Medical Sciences (Beijing, China) and incubated in Dulbecco's modified Eagle's medium (DMEM) containing 10% foetal bovine serum (FBS). After labelling with mouse anti‐human CD90‐fluorescein isothiocyanate (FITC), CD44‐FITC, CD73‐FITC, CD105‐FITC, CD34‐PE, CD14‐PE, CD45‐PC7 and HLA‐DR (all purchased from Abcam Inc, Cambridge, MA, USA), the BM‐MSCs were analysed by flow cytometry. Then, the BM‐MSCs were identified using OriCellTM assay kits (Cyageb Biosciences, Guangzhou, Guangdong, China) by Alizarin Red staining and Oil Red O staining to determine their abilities to differentiate into osteoblasts and adipocytes.