Immunophenotypic Analysis of hAdMSCs

Analyses of the immunophenotype of hAdMSCs were conducted as previously described [19 (link), 20 (link)]. After being washed, hAdMSCs were subjected to incubation for half an hour at 4 °C with primary antibodies against human, including CD34-PE (cat#550761, BD Biosciences), CD44-PE (cat# 559942, BD Biosciences), CD29-PE (cat#556049, BD Biosciences), CD73-PE (cat# 550257, BD Biosciences), CD90-PE (cat# 555596, BD Biosciences), CD45-PE (cat# 560975, BD Biosciences), CD106-PE (cat# 561679, BD Biosciences), CD105-PE (cat# 560839, BD Biosciences), CD206-FITC (cat#5 51135, BD Biosciences) and HLA-DR-PE(cat# 555561, BD Biosciences). Following washing, hAdMSCs were subjected to incubation with secondary antibodies for half an hour at 4 °C. MSCs were then examined using FACSCalibur (BD Biosciences) and the FlowJo program (FlowJo, LCC). Moreover, adipogenic and osteogenic differentiation media were used to cultivate the cells. We performed oil red O staining to assess the adipogenic differentiation of cells (Solarbio, China). Alkaline phosphatase (ALP), an early osteogenic marker, and Alizarin Red (ARS), a late osteogenic marker, were used to quantify the osteogenic differentiation of the cells.