Document Detail

Evaluation of eluents from separations of CD34+ cells from human cord blood using a commercial, immunomagnetic cell separation system.
MedLine Citation:
PMID:  11587583     Owner:  NLM     Status:  MEDLINE    
Human CD34+ cells from cord blood were separated in a two-step process using a commercial, immunomagnetic cell retention system. The performance of the system was evaluated by analyzing a number of eluents from the separations with a number of analytical techniques. In addition to cell counts and flow cytometry analysis, a new experimental technique that is undergoing development, cell tracking velocimetry (CTV), was used. CTV measures the degree to which a cell is immunomagnetically labeled, known as the magnetophoretic mobility, of a population of cells on a cell-by-cell basis and presents the results in the form of a histogram similar to flow cytometry data. The average recovery and purity of CD34+ cells from 10 separations was 52% and 60%, respectively. CTV analysis indicated that the mean magnetophoretic mobility of the positively enriched CD34 cells was 9.64 x 10(-5) mm3/T-A-s, while the mean mobility from negative eluents was -2.02 x 10(-6) mm3/T-A-s, very similar to the mobility of unlabeled cells. Within the positive eluents, the range of magnetophoretic mobility was approximately 50-fold, representing a plausible 50-fold range in surface CD34 antigen expression. CTV analysis also indicated that in some separations, positive cells were not retained by the immunomagnetic cell retention system. Finally, preliminary studies indicate that monocytes might be a primary cause in the lower purities and recoveries seen in this study. It is suggested that the monocytes phagocytose the magnetic nanobeads and become sufficiently magnetized to be retained within the Miltenyi column, reducing the purity of the positive eluent.
K Melnik; M Nakamura; K Comella; L C Lasky; M Zborowski; J J Chalmers
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biotechnology progress     Volume:  17     ISSN:  8756-7938     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2001 Sep-Oct
Date Detail:
Created Date:  2001-10-05     Completed Date:  2002-04-01     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  907-16     Citation Subset:  IM    
Department of Pathology, Ohio State University, 140 West 19th Avenue, Columbus, Ohio 43220, USA.
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MeSH Terms
Antigens, CD34 / analysis*,  immunology
Cell Count
Fetal Blood / cytology
Flow Cytometry
Hematopoietic Stem Cells / cytology*,  immunology
Immunohistochemistry / methods
Immunomagnetic Separation / instrumentation,  methods*,  standards
Grant Support
CA16058-25/CA/NCI NIH HHS; R01 CA62349/CA/NCI NIH HHS; R33 CA81662-01/CA/NCI NIH HHS
Reg. No./Substance:
0/Antigens, CD34

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