title: Micropallet arrays for the capture, isolation and culture of circulating tumor cells from whole blood of mice engrafted with primary human pancreatic adenocarcinoma.
abstract: Circulating tumor cells (CTCs) are important biomarkers of cancer progression and metastatic potential. The rarity of CTCs in peripheral blood has driven the development of technologies to isolate these tumor cells with high specificity; however, there are limited techniques available for isolating target CTCs following enumeration. A strategy is described to capture and isolate viable tumor cells from whole blood using an array of releasable microstructures termed micropallets. Specific capture of nucleated cells or cells expressing epithelial cell adhesion molecules (EpCAM) was achieved by functionalizing micropallet surfaces with either fibronectin, Matrigel or anti-EpCAM antibody. Surface grafting of poly(acrylic acid) followed by covalent binding of protein A/G enabled efficient capture of EpCAM antibody on the micropallet surface. MCF-7 cells, a human breast adenocarcinoma, were retained on the array surface with 90±8% efficiency when using an anti-EpCAM-coated array. To demonstrate the efficiency of tumor cell retention on micropallet arrays in the presence of blood, MCF-7 cells were mixed into whole blood and added to small arrays (71 mm(2)) coated with fibronectin, Matrigel or anti-EpCAM. These approaches achieved MCF-7 cell capture from ≤10 µL of whole blood with efficiencies greater than 85%. Furthermore, MCF-7 cells intermixed with 1 mL blood and loaded onto large arrays (7171 mm(2)) were captured with high efficiencies (≥97%), could be isolated from the array by a laser-based approach and were demonstrated to yield a high rate of colony formation (≥85%) after removal from the array. Clinical utility of this technology was shown through the capture, isolation and successful culture of CTCs from the blood of mice engrafted with primary human pancreatic tumors. Direct capture and isolation of living tumor cells from blood followed by analysis or culture will be a valuable tool for cancer cell characterization.
authors: Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, United States.
authors: Philip C
authors: PC
authors: Gach
authors: Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, NC 27599, United States; Joint Department of Biomedical Engineering, North Carolina State University, Raleigh, NC 27695, United States.
authors: Peter J
authors: PJ
authors: Attayek
authors: Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599, United States.
authors: Rebecca L
authors: RL
authors: Whittlesey
authors: Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599, United States; Department of Surgery, Division of surgical Oncology, University of North Carolina, Chapel Hill, NC 27599, United States; Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599, United States.
authors: Jen Jen
authors: JJ
authors: Yeh
authors: Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, United States; Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, NC 27599, United States; Joint Department of Biomedical Engineering, North Carolina State University, Raleigh, NC 27695, United States; Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599, United States; Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599, United States. Electronic address: nlallbri@unc.edu.
authors: Nancy L
authors: NL
authors: Allbritton
mesh_terms: D000230:Adenocarcinoma / Q000097:blood* / Q000473:pathology; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation*; D004867:Equipment Design; D005260:Female; D006801:Humans; D051379:Mice; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000097:blood* / Q000473:pathology; D046888:Tissue Array Analysis / Q000295:instrumentation*
keywords: Cell array; Cell sorting; Circulating tumor cells; Microfabrication; Pancreatic cancer; Surface functionalization