Cd45 and cd140b

The single cell suspension obtained as described above was stained for cell sorting with a cocktail of biotinylated antibodies for lineage markers, anti-CD2, -CD3, -CD16, -CD64 (BD Biosciences; cat# 555325, 555338, 555405, and 555526), -CD31 (Invitrogen; cat# MHCD3115), -CD45 and -CD140b (BioLegend; cat# 304003 and 323604) to specifically remove hematopoietic, endothelial, and leukocyte lineage cells. Cells were stained for 20 min at room temperature in PBS with 1% BSA, followed by washing in PBS with 1% BSA. Biotinylated primary antibodies were revealed with an anti-human secondary antibody labeled with streptavidin-Pacific Blue conjugate (Invitrogen; cat# S11222). The lineage-positive and negative cell populations were sorted using a FACSAria II cell sorter (BD Biosciences). The lineage-positive population was discarded. The lineage-negative cell population was incubated simultaneously with three human-specific primary antibodies (anti CD-10 labeled with Phycoerythrin (PE) coupled to Cy7 (BD Biosciences; cat# 341092), anti-MUC1 labeled with FITC (BD Biosciences; cat# 559774) and anti-CD73 labeled with PE (phycoerythrin) (BD Biosciences; cat# 550257)) for 20 min at room temperature in PBS with 1% BSA, followed by washing in PBS with 1% BSA. Cells were then sorted as described above.