Isolation of a highly clonogenic and multipotential subfraction of adult stem cells from bone marrow stroma.
Attempts have been made to develop cell and gene therapies using the adult stem cells from bone marrow referred to as mesenchymal stem cells or marrow stromal cells (MSCs). However, the results have been variable in part because there are no standardized protocols for preparing and characterizing MSCs. In the experiments presented here, we developed a standardized assay by light scattering to measure the content of rapidly self-renewing cells (RS cells) in preparations of MSCs. The assay quickly identifies preparations of MSCs that replicate rapidly in subsequent culture. In addition, the standardized assay enabled us to isolate RS cells that were up to 90% clonogenic and that generated single cell-derived colonies that differentiated into either mineralizing cells or adipocytes with appropriate additions to the medium.
Smith
Jason R
JR
Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112, USA.
Pochampally
Radhika
R
Perry
Anthony
A
Hsu
Shu-Ching
SC
Prockop
Darwin J
DJ
D017667:Adipocytes / Q000166:cytology / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D016026:Bone Marrow Transplantation / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D019070:Cell Lineage / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D003114:Colony-Forming Units Assay / Q000379:methods*; D003470:Culture Media / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D039902:Multipotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D010006:Osteoblasts / Q000166:cytology / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D033581:Stem Cell Transplantation / Q000379:methods; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*
D003470:Culture Media; D012333:RNA, Messenger