Anti cd31 pe cy7 conjugated

To identify MPCs and MSC-like cells (Keating, 2012 (link)) in MPC primary cultures freshly detached cells were incubated with anti-CD90 FITC-conjugated, anti-CD73 PE-conjugated, anti-CD31 PE/Cy7-conjugated, anti-CD18 APC-conjugated, and anti-CD45 VioBlue® -conjugated (Miltenyi Biotec, BergischGladbach, Germany) antibodies for 30′ at 4°C in the dark and washed twice in MACSQuant® Running Buffer (Miltenyi Biotec). Data were acquired using MACSQuant® flow cytometer and analyzed by MACSQuantify® Analysis Software (MiltenyiBiotec). MPCs were identified as CD31⁺CD18⁺CD45^lowCD73^negCD90^neg events and MSCs as CD31^negCD18^negCD45^negCD73^brightCD90^bright events. Statistical analysis was performed by one-way analysis of variance (ANOVA) test and Dunnett's post-test for multiple comparison. Results were expressed as mean value ± standard error (SE).

3 × 10⁵ freshly isolated MPCs were processed for immunophenotyping and incubated with anti-CD11c VioBlue®-conjugated, anti-CD18 PE-conjugated, anti-CD31 PE/Cy7-conjugated and anti-CD90 APC-conjugated (all from Miltenyi Biotec, Bergisch Gladbach, Germany) for 30′ at 4°C in the dark. Cells were then washed with MACSQuant® Running Buffer (Miltenyi Biotec) and resuspended in 500 μl of the same buffer for data acquisition in MACSQuant® Flow Cytometer (Miltenyi Biotec). Data were acquired and analyzed by MACSQuantify® analysis software (Miltenyi Biotec). Samples with percentage of CD11c⁺CD18⁺CD31⁺CD90^neg lower than 95% were excluded from the study.