Cd45 allophycoerythrin apc

Single cell suspensions were prepared from normal pregnant women and GDM patient-derived HUCPVC (hereafter called N-PVC and GDM-PVC) cultures by dissociation with 0.05% trypsin-EDTA. The cells were resuspended in 1% FBS-PBS and were stained for 1 hr with the following fluorochrome-conjugated anti-human antibodies: CD31-phycoerythrin (PE), CD34-luorescein-isothiocyanate (FITC), CD45-allophycoerythrin (APC), SSEA-4-FITC, CD146-FTIC, CD44-APC and CD90-APC (all BD Biosciences) or their corresponding isotype controls. After immunostaining, the cells were incubated with the viability dye 7-aminoactinomycin D (7AAD) to exclude dead cells. Data was analyzed using FACSCanto II (BD Biosciences) and FlowJo software.

HUCPVCs were dissociated into single cells using trypsin-EDTA and resuspended in
1% FBS-PBS. The cells were filtered through a 70 µm cell strainer and reacted
with the following antibodies conjugated with fluorochrome for 1 hr at 4℃:
CD31-phycoerythrin (PE), CD34-fluorescein-isothiocyanate (FITC),
CD45-allophycoerythrin (APC), CD44-APC, CD90-APC, CD146-FITC, and SSEA-4-FITC
(all, BD Biosciences). The dead cells were excluded with 7-aminoactinomycin D.
Stained cells were analyzed using the FACSCanto II (BD Biosciences) and data
were analyzed by FlowJo software (FlowJo).