EBs were collected by rinsing with DPBS and treated with 0.05% trypsin at 37°C for 5 min. After digestion, cells were collected and centrifuged at 0.3 relative centrifugal force for 5 min. The pellet was resuspended in flow buffer containing 1% bovine serum in DPBS and incubated for 15 min on ice with 1% fetal bovine serum in DPBS containing the following antibodies at a 1:100 dilution: PE anti-human TNAP Antibody (BioLegend 327305) and FITC anti-human CD38 Antibody (BioLegend 303503). Cells were then washed thrice with the flow buffer, and the resulting single cell suspension was passed through a 40 μm filter and analyzed using a NovoCyte cytometer. Seven-amino-actinomycin D Viability Staining Solution (Thermo Fisher Scientific 00-6993-50) was used as a marker of viability.

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