Document Detail


Characterization of cell seeding and specific capture of B cells in microbubble well arrays.
MedLine Citation:
PMID:  23358874     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Development of micro-well array systems for use in high-throughput screening of rare cells requires a detailed understanding of the factors that impact the specific capture of cells in wells and the distribution statistics of the number of cells deposited into wells. In this study we investigate the development of microbubble (MB) well array technology for sorting antigen-specific B-cells. Using Poisson statistics we delineate the important role that the fractional area of MB well opening and the cell seeding density have on determining cell seeding distribution in wells. The unique architecture of the MB well hinders captured cells from escaping the well and provides a unique microenvironmental niche that enables media changes as needed for extended cell culture. Using cell lines and primary B and T cells isolated from human peripheral blood we demonstrate the use of affinity capture agents coated in the MB wells to enrich for the selective capture of B cells. Important differences were noted in the efficacy of bovine serum albumin to block the nonspecific adsorption of primary cells relative to cell lines as well as the efficacy of the capture coatings using mixed primary B and T cells samples. These results emphasize the importance of using primary cells in technology development and suggest the need to utilize B cell capture agents that are insensitive to cell activation.
Authors:
Meghan C Jones; James J Kobie; Lisa A Delouise
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Biomedical microdevices     Volume:  15     ISSN:  1572-8781     ISO Abbreviation:  Biomed Microdevices     Publication Date:  2013 Jun 
Date Detail:
Created Date:  2013-05-10     Completed Date:  2014-01-13     Revised Date:  2014-08-04    
Medline Journal Info:
Nlm Unique ID:  100887374     Medline TA:  Biomed Microdevices     Country:  United States    
Other Details:
Languages:  eng     Pagination:  453-63     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
B-Lymphocytes / cytology*
Cell Line, Tumor
Cell Separation / instrumentation*
Humans
Microbubbles*
T-Lymphocytes / cytology
Grant Support
ID/Acronym/Agency:
P30 AI078498/AI/NIAID NIH HHS; P30AI078498/AI/NIAID NIH HHS; U19AI056390/AI/NIAID NIH HHS; UL1 TR000038/TR/NCATS NIH HHS; UL1 TR000038/TR/NCATS NIH HHS; UL1 TR000042/TR/NCATS NIH HHS
Comments/Corrections

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