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Initial clinical trials examining the transplantation of ex vivo expanded cord blood (CB) cells have failed to demonstrate an impact on hematopoietic recovery compared with historical unmanipulated CB controls. In this study, we tested whether coculture with primary human brain endothelial cells (HUBECs) could increase the engraftment capacity and repopulating cell frequency within CB CD34 +  cells. Quantitative analysis demonstrated that HUBEC coculture for 7 days supported a 19‐fold greater number of CD34 +  cells and 3.4‐fold and 2.6‐fold greater severe combined immunodeficient (SCID)‐repopulating cell (SRC) frequencies than fresh CB CD34 +  cells and liquid suspension‐cultured cells. Mice transplanted with day‐14 HUBEC‐cultured cells showed 4.2‐fold higher levels of human engraftment than mice transplanted with day‐7 HUBEC‐cultured cells, indicating that SRC enrichment continued to occur through day 14. Noncontact HUBEC cultures also maintained SRCs at levels comparable with contact HUBEC cultures, demonstrating that HUBEC‐secreted soluble factors critically supported SRC self‐renewal. Seeding efficiency studies demonstrated that HUBEC‐cultured CB CD34 +  cells engrafted nonobese diabetic/SCID marrow at significantly higher levels than either fresh CB CD34 +  cells or liquid suspension‐cultured CD34 +  cells. These studies indicate that the application of HUBEC coculture or HUBEC‐conditioned media can potentially improve upon current strategies for the clinical expansion of CB stem cells.

stem cells

Quantitative Analysis Demonstrates Expansion of SCID‐Repopulating Cells and Increased Engraftment Capacity in Human Cord Blood Following Ex Vivo Culture with Human Brain Endothelial Cells