Micropallet arrays with poly(ethylene glycol) walls

Wang, Yuli; Salazar, Georgina To’a; Pai, Jeng Hao; Shadpour, Hamed; Sims, Christopher E.; Allbritton, Nancy L.

doi:10.1039/b800286j

Cited by 22 publications

(22 citation statements)

References 25 publications

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“…Functionalization of anti-EpCAM by attachment to PAA-protein A/G exhibited the greatest surface density of anti-EpCAM and so was employed for all subsequent experiments. Replacement of the virtual air walls by a more rugged microstructure boundary such as poly(ethylene glycol) (PEG) walls 24 or polydimethylsiloxane (PDMS) 25 may permit prolonged incubation with the CAN solution and likely improve the quantity of anti-EpCAM immobilization to the array surfaces.…”

Section: Resultsmentioning

confidence: 99%

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

Gach

Attayek

Whittlesey

et al. 2014

Biosensors and Bioelectronics

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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 mm2) 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 mm2) 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.

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Section: Resultsmentioning

confidence: 99%

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

Gach

Attayek

Whittlesey

et al. 2014

Biosensors and Bioelectronics

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“…Small sized (30-50 μm) microparts enable isolation and culture of single cells [12,[14][15][16][18][19][20][21], and large sized (50-1000 μm) microparts enable multiple cell culture [13,14]. Various types of adherent cells can be cultured on the planar microparts: fibroblast cells [12,13,15,16,28,49,50,55,59], hepatic cells [12,15], muscle cells [13,27,52], neural cells [14,15], cervical cancer cells [15, 18-21, 28, 32, 34], and embryonic stem cells [23,30].…”

Section: Cell Culturementioning

confidence: 99%

Fabrication of 3D Cellular Tissue Utilizing MEMS Technologies

Yoshida

Serien

Tomoike

et al. 2015

Hyper Bio Assembler for 3D Cellular Systems

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This chapter focuses on the use of micro-sized cell culture substrates fabricated by micro electro mechanical systems (MEMS) technologies in the field of three-dimensional (3D) cellular tissue constructions. First, we provide a brief overview of MEMS-based tissue engineering methods, and explain why the micro-sized cell-laden substrates are important. Second, we explain the fabrication processes of the micro-sized cell-laden substrates. The fabrication of micro-sized substrates is described by focusing on their materials, structures, cell culture processes, and handling. Third, their applications for single cell handling is described by providing information on observation, collection, and arrangement. Finally, assembly of 3D cellular constructs is explained by pick-and-place assembly, self-assembly, and self-folding methods. Key advantages of the micro-sized substrates are the ability of manipulating various types of adherent cells and also spatial control of cells in 3D cellular tissues. The single cell handling ability facilitates the cell-cell interaction analysis in the field of pharmacology, and 3D cellular assembly ability will open the route for fabrication of spatially-controlled heterogeneous 3D cellular tissues in the regenerative medicine field.

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“…However, it should be mentioned that microwell arrays are passive structures, which restricts the possibilities to manipulate trapped cells. Therefore, integration of microwell arrays into microfluidic devices (93) or the development of releasable microwells (94) may provide new opportunities to handle and analyze cells cultured in microwells.…”

Section: Microwell Arrays As Pseudo–3-d Niche Modelsmentioning

confidence: 99%

High-Throughput Methods to Define Complex Stem Cell Niches

Köbel

Lütolf

2010

BioTechniques

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The potential of stem cells in clinics and as a diagnostic tool is still largely unmet, partially due to a lack of in vitro models that efficiently mimic the in vivo stem cell microenvironment-or niche-and thus would allow reproducible propagation of stem cells or their controlled differentiation in vitro. The current methodological challenges in studying and manipulating stem cells have spurred intense development and application of microfabrication and micropatterning technologies in stem cell biology. These approaches can be readily used to dissect the complex molecular interplay of stem cells and their niche and study single-cell behavior in high-throughput. Increased merging of microfabrication with advanced biomaterials technologies may ultimately result in functional artificial niches capable of recapitulating extrinsic stem cell regulation in vitro and on a single-cell level.

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Micropallet arrays with poly(ethylene glycol) walls

Cited by 22 publications

References 25 publications

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

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

Fabrication of 3D Cellular Tissue Utilizing MEMS Technologies

High-Throughput Methods to Define Complex Stem Cell Niches

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