Mesenchymal stem cells secreting angiopoietin-like-5 support efficient expansion of human hematopoietic stem cells without compromising their repopulating potential

Stem Cells Dev. 2011 Aug;20(8):1371-81. doi: 10.1089/scd.2010.0456. Epub 2011 Jan 31.


Clinical and preclinical applications of human hematopoietic stem cells (HSCs) are often limited by scarcity of cells. Expanding human HSCs to increase their numbers while maintaining their stem cell properties has therefore become an important area of research. Here, we report a robust HSC coculture system wherein cord blood CD34(+) CD133(+) cells were cocultured with mesenchymal stem cells engineered to express angiopoietin-like-5 in a defined medium. After 11 days of culture, SCID repopulating cells were expanded ~60-fold by limiting dilution assay in NOD-scid Il2rg(-/-) (NSG) mice. The cultured CD34(+) CD133(+) cells had similar engraftment potential to uncultured CD34(+) CD133(+) cells in competitive repopulation assays and were capable of efficient secondary reconstitution. Further, the expanded cells supported a robust multilineage reconstitution of human blood cells in NSG recipient mice, including a more efficient T-cell reconstitution. These results demonstrate that the expanded CD34(+) CD133(+) cells maintain both short-term and long-term HSC activities. To our knowledge, this ~60-fold expansion of SCID repopulating cells is the best expansion of human HSCs reported to date. Further development of this coculture method for expanding human HSCs for clinical and preclinical applications is therefore warranted.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen
  • Angiopoietin-like Proteins
  • Angiopoietins / biosynthesis*
  • Angiopoietins / genetics
  • Angiopoietins / metabolism
  • Animals
  • Antigens, CD / biosynthesis
  • Antigens, CD34 / biosynthesis
  • Antigens, CD34 / blood
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Fetal Blood / cytology
  • Fetal Blood / transplantation
  • Glycoproteins / biosynthesis
  • Graft Survival / immunology
  • Hematopoietic Stem Cell Transplantation*
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / immunology
  • Hematopoietic Stem Cells / metabolism*
  • Humans
  • Mesenchymal Stem Cell Transplantation
  • Mesenchymal Stem Cells / metabolism*
  • Mice
  • Peptides
  • T-Lymphocytes / transplantation
  • Transplantation, Heterologous / immunology


  • AC133 Antigen
  • ANGPTL5 protein, human
  • Angiopoietin-like Proteins
  • Angiopoietins
  • Antigens, CD
  • Antigens, CD34
  • Glycoproteins
  • PROM1 protein, human
  • Peptides
  • Prom1 protein, mouse