title: Small sample sorting of primary adherent cells by automated micropallet imaging and release.
abstract: Primary patient samples are the gold standard for molecular investigations of tumor biology yet are difficult to acquire, heterogeneous in nature and variable in size. Patient-derived xenografts (PDXs) comprised of primary tumor tissue cultured in host organisms such as nude mice permit the propagation of human tumor samples in an in vivo environment and closely mimic the phenotype and gene expression profile of the primary tumor. Although PDX models reduce the cost and complexity of acquiring sample tissue and permit repeated sampling of the primary tumor, these samples are typically contaminated by immune, blood, and vascular tissues from the host organism while also being limited in size. For very small tissue samples (on the order of 10(3) cells) purification by fluorescence-activated cell sorting (FACS) is not feasible while magnetic activated cell sorting (MACS) of small samples results in very low purity, low yield, and poor viability. We developed a platform for imaging cytometry integrated with micropallet array technology to perform automated cell sorting on very small samples obtained from PDX models of pancreatic and colorectal cancer using antibody staining of EpCAM (CD326) as a selection criteria. These data demonstrate the ability to automate and efficiently separate samples with very low number of cells.
authors.affiliation: Department of Biomedical Engineering, University of North Carolina, Chapel Hill, North Carolina 27599 and North Carolina State University, Raleigh, North Carolina, 27695.
authors.forename: Pavak K
authors.initials: PK
authors.lastname: Shah
authors.forename: Silvia Gabriela
authors.initials: SG
authors.lastname: Herrera-Loeza
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Sims
authors.forename: Jen Jen
authors.initials: JJ
authors.lastname: Yeh
authors.forename: Nancy L
authors.initials: NL
authors.lastname: Allbritton
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis*; D015815:Cell Adhesion Molecules / Q000032:analysis*; D015179:Colorectal Neoplasms / Q000473:pathology*; D001330:Electronic Data Processing; D000071858:Epithelial Cell Adhesion Molecule; D005434:Flow Cytometry / Q000379:methods*; D064593:Heterografts / Q000166:cytology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008819:Mice, Nude; D010190:Pancreatic Neoplasms / Q000473:pathology*; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule
keywords: cell sorting; image cytometry; micropallet arrays; xenografts