Cord Blood Lin2CD452 Embryonic-Like Stem Cells Are a Heterogeneous Population That Lack Self-Renewal Capacity

Human umbilical cord blood (hUCB) has been proposed to contain not only haematopoietic stem cells, but also a rare pluripotent embryonic-like stem cell (ELSc) population that is negative for hematopoietic markers (Lin2CD452) and expresses markers typical of pluripotent cells. The aim of this work was to isolate, characterise and expand this ELSc fraction from hUCB, as it may provide a valuable cell source for regenerative medicine applications. We found that we could indeed isolate a Lin2CD452 population of small cells (3–10 mm diameter) with a high nucleus to cytoplasm ratio that expressed the stem cell markers CD34 and CXCR4. However, in contrast to some previous reports, this fraction was not positive for CD133. Furthermore, although these cells expressed transcripts typical of pluripotent cells, such as SOX2, OCT3/4, and NANOG, they were not able to proliferate in any of the culture media known to support stem cell growth that we tested. Further analysis of the Lin2CD452 population by flow cytometry showed the presence of a Lin2CD452Nestin+ population that were also positive for CD34 (20%) but negative for CXCR4. These data suggest that the Lin2CD452 stem cell fraction present in the cord blood represents a small heterogeneous population with phenotypic characteristics of stem cells, including a Lin2CD452Nestin+ population not previously described. This study also suggests that heterogeneity within the Lin2CD452 cell fraction is the likely explanation for differences in the hUCB cell populations described by different groups that were isolated using different methods. These populations have been widely called ‘‘embryonic-like stem cell’ ’ on the basis of thei