Details

Autor(en) / Beteiligte

• Gach, Philip C.
• Attayek, Peter J.
• Whittlesey, Rebecca L.
• Yeh, Jen Jen
• Allbritton, Nancy L.

Titel

Micropallet arrays for the capture, isolation and culture of circulating tumor cells from whole blood of mice engrafted with primary human pancreatic adenocarcinoma

Ist Teil von

• Biosensors & bioelectronics, 2014-04, Vol.54, p.476-483

Ort / Verlag

Kidlington: Elsevier B.V

Erscheinungsjahr

2014

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 (71mm2) coated with fibronectin, Matrigel or anti-EpCAM. These approaches achieved MCF-7 cell capture from ≤10µL of whole blood with efficiencies greater than 85%. Furthermore, MCF-7 cells intermixed with 1mL blood and loaded onto large arrays (7171mm2) 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. •CTCs were captured from whole blood on an array.•The array elements were releasable enabling viable CTC isolation.•Greater than 85% of tumor cells were captured from up to 1mL of whole blood.•CTCs were captured and cultured from a xenograft model of human pancreatic adenocarcinoma.