After 21 days of incubation with CGF at serum-reduced conditions, multipotency of ADSC was assessed via flow cytometry analysis of cell surface protein markers, using the CytoFLEX LX flow cytometer (Beckman Coulter, CA, United States). Briefly, ADSC was first harvested using Accutase solution and resuspended in 0.1% BSA in 1× PBS at the cell density of 1 × 106 cells/mL. The cells were then stained with FITC anti-human CD105 (Clone MEM-229), CD73 (Clone AD2), and CD90 (Clone 5E10), as well as PE/Cy5.5 anti-human CD45 (all Abcam, Cambridge, United Kingdom), at 4°C for 30 min in the dark, according to the manufacturer’s protocol. Untreated and unstained ADSC was assessed in the same manner for the quadrants to be drawn. Percentage of positive cells for each marker was calculated from three independent experiments (n = 3).
Pe cy5.5 anti human cd45
Manufactured by Abcam
Sourced in United Kingdom
PE/Cy5.5 anti-human CD45 is a fluorescently-labeled antibody that binds to the CD45 surface antigen on human cells. CD45 is a protein tyrosine phosphatase that is expressed on all nucleated hematopoietic cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pe cy5.5 anti human cd45
1
Multipotency Characterization of ADSC
2
Characterizing Cryopreserved ADSC Phenotype
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After 14 days
of culture, the cell phenotype of encapsulated ADSCs
cryopreserved with 0.75 M trehalose was assessed via flow cytometry
using a CytoFLEX S flow cytometer (Beckman Coulter, CA, USA). Briefly,
ADSCs were first harvested using the Accutase solution and resuspended
in 1× PBS at a cell density of 1 × 106 cells/mL.
The cell suspension was then divided into three equal portions and
separately stained with FITC anti-human CD105 (Clone MEM-229), CD73
(Clone AD2), or CD90 (Clone 5E10). Next, all the cells were concurrently
stained with secondary PE/Cy5.5 anti-human CD45 (Clone HI30) (all
from Abcam, Cambridge, UK). Cells were stained at 4 °C for 15
min in the dark according to the manufacturer’s protocol. Untreated
and unstained ADSCs expressing negligible fluorescence were assessed
in the same manner for the quadrants to be drawn. The percentage of
positive cells for each marker was averaged from three independent
experiments (n = 3).
of culture, the cell phenotype of encapsulated ADSCs
cryopreserved with 0.75 M trehalose was assessed via flow cytometry
using a CytoFLEX S flow cytometer (Beckman Coulter, CA, USA). Briefly,
ADSCs were first harvested using the Accutase solution and resuspended
in 1× PBS at a cell density of 1 × 106 cells/mL.
The cell suspension was then divided into three equal portions and
separately stained with FITC anti-human CD105 (Clone MEM-229), CD73
(Clone AD2), or CD90 (Clone 5E10). Next, all the cells were concurrently
stained with secondary PE/Cy5.5 anti-human CD45 (Clone HI30) (all
from Abcam, Cambridge, UK). Cells were stained at 4 °C for 15
min in the dark according to the manufacturer’s protocol. Untreated
and unstained ADSCs expressing negligible fluorescence were assessed
in the same manner for the quadrants to be drawn. The percentage of
positive cells for each marker was averaged from three independent
experiments (n = 3).
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