Document Detail

Isolation and culture of bone marrow-derived human multipotent stromal cells (hMSCs).
MedLine Citation:
PMID:  18370080     Owner:  NLM     Status:  MEDLINE    
We have developed protocols whereby a total of 30-90 x 10(6) hMSCs with an average viability greater than 90% can be produced in a single multilevel Cell Factory from a relatively small (1-3 mL) bone marrow aspirate in 14-20 d. It is possible to generate as many as 5 x 10(8) multipotent stromal cells (MSCs) from a single sample, depending on the number of Cell Factories seeded from the initial isolated hMSCs. Briefly, mononuclear cells are collected from a bone marrow aspirate by density gradient centrifugation. The cells are cultured overnight and the adherent cells are allowed to attach to the flask. Nonadherent cells are removed and the culture expanded for 7-10 d with periodic feeding of the cells. The cells are then harvested and seeded at low density (60-100 cells/cm2) into Nunc Cell Factories. The cells are allowed to expand for an additional 7-10 d, and are then harvested.
Margaret Wolfe; Radhika Pochampally; William Swaney; Roxanne L Reger
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  449     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2008  
Date Detail:
Created Date:  2008-03-28     Completed Date:  2008-06-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3-25     Citation Subset:  IM    
Center for Gene Therapy, Tulane University Health Sciences Center, New Orleans, LA, USA.
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MeSH Terms
Bone Marrow Cells / cytology*
Cell Culture Techniques / methods*
Cells, Cultured
Mesenchymal Stem Cells / cytology*
Multipotent Stem Cells / cytology*
Stromal Cells / cytology

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