Adhesion to fibronectin maintains regenerative capacity during ex vivo culture and transduction of human hematopoietic stem and progenitor cells.
Recent reports have indicated that there is poor engraftment from hematopoietic stem cells (HSC) that have traversed cell cycle ex vivo. However, inducing cells to cycle in culture is critical to the fields of ex vivo stem cell expansion and retroviral-mediated gene therapy. Through the use of a xenograft model, the current data shows that human hematopoietic stem and progenitor cells can traverse M phase ex vivo, integrate retroviral vectors, engraft, and sustain long-term hematopoiesis only if they have had the opportunity to engage their integrin receptors to fibronectin during the culture period. If cultured in suspension under the same conditions, transduction is undetectable and the long-term multilineage regenerative capacity of the primitive cells is severely diminished.
Dao
M A
MA
Division of Research Immunology/Bone Marrow Transplantation, Childrens Hospital Los Angeles, CA, USA.
Hashino
K
K
Kato
I
I
Nolta
J A
JA
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002999:Clone Cells / Q000737:chemistry / Q000166:cytology; D003114:Colony-Forming Units Assay; D004351:Drug Resistance / Q000235:genetics; D005353:Fibronectins / Q000378:metabolism*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics / Q000494:pharmacology; D008565:Membrane Proteins / Q000494:pharmacology; D051379:Mice; D008819:Mice, Nude; D017154:Stromal Cells / Q000378:metabolism / Q000637:transplantation; D014018:Tissue Distribution; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous
D005353:Fibronectins; D007377:Interleukin-3; D008565:Membrane Proteins; C084881:flt3 ligand protein