PSC are anchored within the niche by expression of the surface marker
CD49f or integrin a6, which, when paired with either [beta]1 or [beta]4, can form an integrin receptor that binds to its ligand laminin, a protein found in its basement membrane and extracellular matrix (ECM) (53,57).
HeLa MonoLayer was cultured for 48 hours, and HeLa SPheres for 7 days for
CD49f detection.
The single-cell suspension obtained was then incubated with 5% BSA (Sigma) in PBS for 30 minutes at 4C0, washed with 0.1% BSA in PBS, and incubated with an Alexa Fluor 488 1:100 conjugated antibody against the
CD49f surface marker (Biolegend) on ice 1h.
CD105, CD44, CD29, 9 (CD9),
CD49f, CD90, major histocompatibility complex class I (MHC I), multipotent markers octamer-binding transcription factor 4 (Oct-4; the main regulator of differentiation in pluripotent cell line), stage-specific embryonic antigen 4 (SSEA-4; the marker used to differentiate between human and mouse cells and used to characterize pluripotent stem cells), and stromal-derived factor-1 (STRO1; used as identification, isolation, and functional testing of human bone marrow stromal progenitor cells) have been identified under fluorescent microscopy or by flow cytometry.
The cells consistently differentiated into three lineages in culture, were negative for hematopoietic markers (CD34, CD36, CD117, and CD45), and were positive for mesenchymal markers CD29 (95%), CD44 (>93%), CD49c (99%),
CD49f (>70%), CD59 (>99%), CD90 (>99%), CD105 (>99%), and CD166 (>99%).
Machida and colleagues (2012) also found that TISCs isolated from the livers of alcohol-fed HCV Ns5a transgenic mice and from alcoholic patients carried molecules called CD133 and
CD49f (i.e., were CD[133.sup.+]/CD[49f.sup.+] cells).
303004; Biolegend), Alexa 488 conjugated-Rat IgG2a against integrin [alpha]6 (
CD49f; cat.