(20,) (21) The expression of the surface marker of the stem cells was analyzed via the incubation of at least 100 000 cells with fluorescence-labeled monoclonal antibodies against
CD29, CD34, CD44, CD45, and CD90 (Sigma, China).
The gene of Oct-4, Rex-1,
CD29, CD44, CD71, and CD73 were identified by RT-PCR and immunofluorescence technique.
Results: ADSCs were successfully isolated from rat adipose tissue as identified by expression of typical surface markers
CD29, CD44, CD90, and CD45 in flow cytometry.
MenSCs and BMSCs expressed MSCs markers including CD9, CD10,
CD29, CD44, CD73 and CD105, they were also negative for hematopoietic markers, CD34, CD38, CD45 and CD133.
Subsequently, all MSCs at passage 4 homogeneously showed positive expression for mesenchymal markers
CD29 (ADSC, 99.53% [+ or -] 0.12%; UCMSC, 98.7%0 [+ or -] 0.30%; EMSC, 90.75% [+ or -] 0.05%), CD90 (ADSC, 97.60% [+ or -] 1.21%; UCMSC, 98.4% [+ or -] 0.33%; EMSC, 92.55% [+ or -] 0.05%), CD73 (ADSC, 98.75% [+ or -] 0.85%; UCMSC, 97.9% [+ or -] 0.5%; EMSC, 99.8% [+ or -] 0.2%), and CD105 (ADSC, 98.95% [+ or -] 0.45%; UCMSC, 93.4% [+ or -] 1.02%; EMSC, 96.95% [+ or -] 0.45%) and were negative for CD31 (<1%), CD34 (<2%), CD45 (<2%), CD14 (<1%), CD19 (<1%), and HLA-DR (<1%)
Third-passage DFSCs were incubated with the antibodies of positive markers for MSCs of human
CD29 allophycocyanin (APC), CD73 phycoerythrin (PE), CD105 PE, and CD146 fluorescein isothiocyanate (FITC) and with the antibodies of negative markers of CD14 PE, CD34 FITC, CD45 APC, and HLA-DR PE (BD Biosciences, USA) for 15 min at room temperature in the dark.
The cells presented high positivity for the following expressions:
CD29 (98.2 [+ or -] 1.1%), CD44 (76.65 [+ or -] 2.55%), CD73 (83.2 [+ or -] 15%), and CD90 (90.05 [+ or -] 8.65%).