Differentiation of human CD34+CD38- cord blood stem cells into B cell progenitors in vitro

Umbilical cord blood CD34+CD38- cells are a rare, quiescent, primitive progenitor subpopulation lacking expression of lymphoid and myeloid lineage specific antigens. Although myeloid, erythroid, and megakaryocytic differentiation from these cells has been described, B lineage differentiation has not been demonstrated previously. We report here that highly enriched human B cell progenitors can be consistently generated from CD34+CD38- cord blood cells using long-term culture on the murine stromal line, S17, in the absence of added cytokines. After 6-8 weeks, cell numbers increased up to 160-fold, and cultures contained > 80-90% CD10+CD19+ B progenitors. Consistent with previous reports describing delayed myeloid cell differentiation from CD34+CD38- cells, production of B cell progenitors from CD34+CD38- cord blood cells was delayed 2-4 weeks relative to cultures initiated with either CD34+CD38bright or CD34+CD38dim progenitors. Addition of Flt3 ligand to long-term cultures resulted in a 2-3-fold greater increase in the number of CD19+ cells generated after 4-8 weeks. The selective outgrowth of B cell progenitors using this culture model will be useful for a range of in vitro studies using primitive hematopoietic progenitors.