CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages

The CD34 antigen serves as an important marker for primitive hematopoietic cells in therapeutic transplantation of hematopoietic stem cells (HSC) and gene therapy, but it has remained an open question as to whether or not most HSC express CD34. Using a competitive long-term reconstitution assay, the results of this study confirm developmental changes in CD34 expression on murine HSC. In fetuses and neonates, CD34 was expressed on Lin^-c-Kit⁺ long-term repopulating HSC of bone marrow (BM), liver, and spleen. However, CD34 expression on HSC decreased with aging, and in mice older than 10 weeks, HSC were most enriched in the Lin^-c-Kit⁺CD34^- marrow cell fraction. A second transplantation was performed from primary recipients who were transplanted with neonatal Lin^-c-Kit⁺ CD34^high HSC marrow. Although donor-type HSC resided in CD34-Expressing cell fraction in BM cells of the first recipients 4 weeks after the first transplantation, the stem cell activity had shifted to Lin^-c-Kit⁺CD34^- cells after 16 weeks, indicating that adult Lin^-c-Kit⁺CD34^- HSC are the progeny of neonatal CD34-expresssing HSC. Assays for colony-forming cells showed that hematopoietic progenitor cells, unlike HSC, continue to express CD34 throughout murine development. The present findings are important because the clinical application of HSC can be extended, in particular as related to CD34-Enriched HSC and umbilical cord blood HSC.