Enrichment and expansion of cells using antibody-coated micropallet arrays.
Positive selection, sorting, and collection of single cells from within a heterogeneous population are required for many biological studies. We recently demonstrated a miniaturized cell array for this purpose; however, on-chip pre-enrichment and isolation of specific target cells would provide significant value for cell isolation. In the current work, mixed cell samples of fewer than 30,000 cells were used for panning by means of on-array antibody-capture to pre-enrich the target population. The cell surface receptors Fc(epsilon)R(1), c-Kit, and ErbB2 were used for positive selection of RBL, RBL, and SK-BR-3 cells, respectively, from the mixed population. The capture efficiency, selectivity, and enrichment for the target cells were calculated and compared with fibronectin-coated controls. As expected, the capture efficiency depended on the frequency of the target cell in the mixed population over the range of 0.3-33%. For a frequency of 5% target cells, the capture efficiency was 39%-53% for the three conditions, while the selectivity varied between 78% and 98% with 16-20-fold enrichment. Furthermore, single-cell cloning studies demonstrated a high cloning efficiency of target cells selectively isolated from the array. Antibody-based pre-enrichment in combination with micropallet-based cell selection will be a valuable tool for isolation and expansion of rare cells from small heterogeneous populations.
Shadpour
Hamed
H
Department of Chemistry, University of North Carolina, Chapel Hill, 27599, USA.
Sims
Christopher E
CE
Allbritton
Nancy L
NL
D000906:Antibodies / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D019009:Proto-Oncogene Proteins c-kit / Q000276:immunology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology; D046888:Tissue Array Analysis / Q000379:methods*
D000906:Antibodies; D019009:Proto-Oncogene Proteins c-kit; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human