By cell surface phenotype and vasculogenic potential in vitro and in vivo, we also found the ECFCs to be extremely similar to AT-ECs.
Cell surface phenotyping was performed by using anti-mouse antibodies to CD3, CD4, CD8, CD19, CD11c, CD11b, MHCII, CD86, CD49b, CD25, CD44, CD138 and/or fixable live/dead.
The development and progression of breast tumours has been proposed to be driven by breast cancer stem cells (CSC) identified by the cell surface phenotype ESA+CD44+CD24low or aldehyde dehydrogenase (ALDH1) activity [ 7, 8].
Different research groups have reported that liver CSC fractions could be successfully enriched by some cell surface phenotypes, specifically CD133, CD90, CD44, epithelial cell adhesion molecule (EpCAM), OV6 and CD13 [ 4- 9].
The cells consisted of >90% mature myeloid DCs as determined by their morphology and their cell surface phenotype (i.e CD1a+, CD80+ and CD86+).
FITC and PE-labeled mAb specific for the DC maturation markers: CD80, CD83, CD86 and HLA-DR (BD Bioscience) and isotype-matched labeled controls were used to characterize cell surface phenotypes by flow cytometry.
FITC- and PE-labelled mAbs against DC markers (BD Biosciences Pharmingen, San Jose, CA, USA) and isotype-matched labelled controls were used to characterise cell surface phenotypes by flow cytometry.
The TCR specificity of clones was validated by FACS using staining with mAbs specific for CD3, Vα24, Vβ11 and the invariant iNKT cell TCR chain (6B11), and the cell surface phenotype was determined by staining with CD4 and CD8α specific mAbs.
Cell surface phenotypes were determined by flow cytometry, on a FACScan flow cytometer (Becton-Dickinson, San Jose, CA).
