For the identification of GA-MSCs, flow cytometry and induced differentiation were performed as we described previously [14 (link)]. In brief, GA-MSCs were collected and stained with fluorochrome-conjugated antibodies, including anti-CD105-APC, anti-CD90-PerCP, anti-CD73-APC/Cy7, anti-CD44-FITC, anti-CD133-APC and anti-CD34-APC (all from eBioscience. USA) in the dark at 4 °C for 30 min. Then, the cells were centrifuged, resuspended and analyzed using a FACS flow cytometer (BD FACSCanto2, Biosciences). The data were collected and analyzed using FlowJo software (v10.6.2). GA-MSCs were differentiated into osteocytes, adipocytes, and chondrocytes by using ready-to-use differentiation media (all from Cyagen, China) following the manufacturer’s instructions. Adipogenic differentiation was evaluated by Oil Red O staining, osteogenic differentiation was evaluated by Alizarin Red staining, and chondrogenic differentiation was evaluated by Alcian Blue staining. The stain results were observed with an inverted phase contrast microscope (Olympus IX73), and photographs were taken with a digital camera using Image-Pro Plus 6.0 software.
Anti cd73 apc cy7
Manufactured by Thermo Fisher Scientific
The Anti-CD73-APC/Cy7 is a fluorochrome-conjugated monoclonal antibody that binds to the CD73 (5'-nucleotidase) cell surface antigen. CD73 is involved in the conversion of extracellular adenosine monophosphate (AMP) to adenosine, which can have immunosuppressive effects. This antibody can be used for the identification and enumeration of CD73-expressing cells in flow cytometry applications.
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Lab products found in correlation
Anti cd105 apc, by Thermo Fisher Scientific (2 mentions)
Anti cd44 fitc, by Thermo Fisher Scientific (1 mentions)
Facs flow cytometer, by BD (1 mentions)
Anti cd34 fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd44 apc cy7, by Thermo Fisher Scientific (1 mentions)
Anti cd14 percp, by Thermo Fisher Scientific (1 mentions)
Anti cd31 pe cy7, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti cd73 apc cy7
1
Identification and Characterization of GA-MSCs
2
Phenotypic Characterization of gb-MSCs
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Flow cytometry analysis was performed using fluorochrome-conjugated antibodies. Briefly, subconfluent gb-MSCs were cultured in flasks with serum-free medium (0% DMEM), standard medium containing 10% FBS (10% DMEM), serum-free glioblastoma-conditioned medium (0% gb-CM) and standard glioblastoma-conditioned medium(s-gb-CM) for 72 h. The cells were trypsinized and collected in PBS. After centrifugation, the resuspended cells were stained with fluorochrome-conjugated antibodies, including anti-CD31-PE/Cy7, anti-CD34-FITC, anti-CD73-APC/Cy7, anti-CD90-PE/Cy7, anti-CD14-percp, anti-CD105-APC and anti-CD44-APC/Cy7 (all from ebioscience. USA) as well as anti-PDGFR-β-PE (R&D, USA) in the dark at 4°C for 30 min. Then, the cells were centrifuged, resuspended in PBS and analyzed using a FACS flow cytometer (BD Biosciences). The data were collected and analyzed using FlowJo (TreeStar, Ashland, OR) software.
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