Endothelial Cells Promote Expansion of Long‐Term Engrafting Marrow Hematopoietic Stem and Progenitor Cells in Primates

• Published in: Stem cells translational medicine 2017-03, Vol.6 (3), p.864-876
• Main Authors: Gori, Jennifer L., Butler, Jason M., Kunar, Balvir, Poulos, Michael G., Ginsberg, Michael, Nolan, Daniel J., Norgaard, Zachary K., Adair, Jennifer E., Rafii, Shahin, Kiem, Hans‐Peter
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD34 - metabolism; Autografts; Bioinformatics; Blood diseases; Bone marrow; Bone Marrow Cells - cytology; Bone marrow transplantation; CD34 antigen; CD34+ cells; Cell Lineage; Cell Proliferation; Cell self-renewal; Colonies; Cytokines; Disease; Editing; Enabling Technologies for Cell‐Based Clinical Translation; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - metabolism; Erythrocytes; Experiments; Gene Expression Profiling; Gene therapy; Genetic disorders; Green fluorescent protein; Hematopoiesis; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Hematopoietic stem progenitor cell; Humans; Leukocytes (granulocytic); Ligands; Lymphocytes; Macrophages; Medical research; Monocytes; Neutrophils; Patients; Polymerase chain reaction; Primates; Progenitor cells; Stem cell transplantation; Stem cells; Studies; Time Factors; Translational s and Reviews
• ISSN: 2157-6564; 2157-6580
• DOI: 10.5966/sctm.2016-0240

Successful expansion of bone marrow (BM) hematopoietic stem and progenitor cells (HSPCs) would benefit many HSPC transplantation and gene therapy/editing applications. However, current expansion technologies have been limited by a loss of multipotency and self‐renewal properties ex vivo. We hypothesized that an ex vivo vascular niche would provide prohematopoietic signals to expand HSPCs while maintaining multipotency and self‐renewal. To test this hypothesis, BM autologous CD34+ cells were expanded in endothelial cell (EC) coculture and transplanted in nonhuman primates. CD34+C38− HSPCs cocultured with ECs expanded up to 17‐fold, with a significant increase in hematopoietic colony‐forming activity compared with cells cultured with cytokines alone (colony‐forming unit‐granulocyte‐erythroid‐macrophage‐monocyte; p < .005). BM CD34+ cells that were transduced with green fluorescent protein lentivirus vector and expanded on ECs engrafted long term with multilineage polyclonal reconstitution. Gene marking was observed in granulocytes, lymphocytes, platelets, and erythrocytes. Whole transcriptome analysis indicated that EC coculture altered the expression profile of 75 genes in the BM CD34+ cells without impeding the long‐term engraftment potential. These findings show that an ex vivo vascular niche is an effective platform for expansion of adult BM HSPCs. Stem Cells Translational Medicine 2017;6:864–876