title: Binding and isolation of tumor cells in biological media with perfluorocarbon microbubbles.
abstract: With the emerging interest in personalized medicine, there is strong demand for new technologies for clinical sample interrogation. Exfoliated tumor cells in variety of pathological samples (e.g., blood, bone marrow, urine) could provide invaluable information for diagnosis and prognosis of cancers. Here we describe a detailed method for capture and isolation of tumor cells in medium, blood, or large issue buffy coat using EpCAM-targeted buoyant microbubbles (MBs). Perflorohexane gas lipid shell MBs were prepared with emulsification method and conjugated with antibody as described by us before [25]. The binding of EpCAM-targeted MBs to A549 (human lung carcinoma) and 4T1 (mouse breast carcinoma) cells spiked into BSA/PBS or blood was more than 90%, which was comparable with commercial anti-EpCAM immunomagnetic beads (DynaBeads). Anti-EpCAM MBs efficiently (75-82%) isolated BxPC3 pancreatic tumor cells spiked into medium, blood or a buffy coat, within 15-30 min of incubation. We discuss MB parameters and experimental conditions critical to achieve efficient cells binding and isolation. In conclusion, MB-assisted cell isolation is a promising method for rapid enrichment of cells and biomarkers from biological samples.
authors: Solid Tumor Therapeutics Program, Moores UCSD Cancer Center, UC San Diego, La Jolla, CA 92093, USA.
authors: Guixin
authors: G
authors: Shi
authors: Wenjin
authors: W
authors: Cui
authors: Rajesh
authors: R
authors: Mukthavaram
authors: Yu-Tsueng
authors: YT
authors: Liu
authors: Dmitri
authors: D
authors: Simberg
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D018914:Cells, Immobilized / Q000166:cytology; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D005466:Fluorocarbons; D006801:Humans; D015674:Mammary Neoplasms, Animal / Q000473:pathology; D051379:Mice; D045423:Microbubbles; D009360:Neoplastic Cells, Circulating / Q000473:pathology*
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D005466:Fluorocarbons
keywords: Blood; CTC; EpCAM; Isolation; Microbubbles
references: Echocardiography. 1999 Oct;16(7, Pt 2):733-741
references: 11175216
references: Nature. 2007 Dec 20;450(7173):1235-9
references: 18097410
references: Clin Cancer Res. 2007 Feb 1;13(3):920-8
references: 17289886
references: Mol Diagn. 2001 Jun;6(2):79-91
references: 11468693
references: J Clin Oncol. 2003 Mar 1;21(5):767-73
references: 12610172
references: Clin Exp Metastasis. 2007;24(5):317-27
references: 17530423
references: Urology. 2001 Sep;58(3):386-92
references: 11549486
references: Clin Cancer Res. 2001 Dec;7(12):4080-5
references: 11751505
references: J Am Chem Soc. 2008 Jul 9;130(27):8633-41
references: 18557614
references: Clin Cancer Res. 2004 May 1;10(9):3020-8
references: 15131038
references: Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10501-4
references: 15249663
references: Lab Chip. 2007 Apr;7(4):463-8
references: 17389962
references: Nat Rev Cancer. 2004 Jun;4(6):448-56
references: 15170447
references: PLoS One. 2013;8(3):e58017
references: 23516425
references: N Engl J Med. 2004 Aug 19;351(8):781-91
references: 15317891
references: Clin Cancer Res. 2000 Sep;6(9):3552-9
references: 10999743
references: Cancer Res. 2010 Aug 15;70(16):6420-6
references: 20663903
references: J Clin Oncol. 2007 Nov 20;25(33):5287-312
references: 17954709
references: Angew Chem Int Ed Engl. 2003 Jul 21;42(28):3218-35
references: 12876730
references: J Surg Oncol. 1991 May;47(1):32-6
references: 2023419
references: Int J Oncol. 2005 Jul;27(1):49-57
references: 15942643
references: Clin Cancer Res. 2003 Oct 15;9(13):4865-71
references: 14581359
references: J Colloid Interface Sci. 2001 Jun 1;238(1):37-42
references: 11350133
references: Clin Cancer Res. 2002 Jul;8(7):2073-84
references: 12114406
references: Langmuir. 2008 Mar 4;24(5):1745-9
references: 18205422
references: Nat Med. 2006 Aug;12(8):895-904
references: 16892035