ADSCs at passage 3 were cultured in osteogenic, adipogenic, and chondrogenic differentiation media (Cyagen Biosciences, Guangzhou, China). After incubation, the ADSCs were fixed with 4% paraformaldehyde and subsequently identified using Alizarin Red, Oil Red O, and Alcian Blue staining.
Cell surface marker detection of ADSCs was performed as previously described [57 (link)]. ADSCs at passage 3 were diluted at a density of 5 × 105 cells/100 µL in a staining buffer consisting of PBS supplemented with 4% FBS. The cells were then incubated with specific antibodies, including anti-CD31, anti-CD34, anti-CD45, anti-CD34, anti-29, anit-CD90, and anti-CD105 (all direct-labeled antibodies from BD Biosciences, Franklin Lakes, NJ, USA). The incubation was conducted at 4 °C for 30 min. After being washed twice, ADSCs were prepared in 100 µL of staining buffer and analyzed by flow cytometry (CytoFLEX LX, Beckman Coulter, Brea, CA, USA).
Cell surface marker detection of ADSCs was performed as previously described [57 (link)]. ADSCs at passage 3 were diluted at a density of 5 × 105 cells/100 µL in a staining buffer consisting of PBS supplemented with 4% FBS. The cells were then incubated with specific antibodies, including anti-CD31, anti-CD34, anti-CD45, anti-CD34, anti-29, anit-CD90, and anti-CD105 (all direct-labeled antibodies from BD Biosciences, Franklin Lakes, NJ, USA). The incubation was conducted at 4 °C for 30 min. After being washed twice, ADSCs were prepared in 100 µL of staining buffer and analyzed by flow cytometry (CytoFLEX LX, Beckman Coulter, Brea, CA, USA).
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