title: A Microfabricated 96-Well 3D Assay Enabling High-Throughput Quantification of Cellular Invasion Capabilities.
abstract: This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation. Invasion of the tumor cells into the extracellular matrix was subsequently initiated and monitored. Two modes of cellular infiltration were observed: A549 cells invaded into the extracellular matrix following the surfaces previously coated with PEG molecules in a pseudo-2D manner, while H1299 cells invaded into the extracellular matrix in a truly 3D manner including multiple directions. Based on the processing of 2D microscopic images, a key parameter, namely, equivalent invasion distance (the area of invaded cells divided by the circumference of the initial cell cluster) was obtained to quantify migration capabilities of these two cell types. These results validate the feasibility of the proposed platform, which may function as a high-throughput 3D cellular invasion assay.
authors.affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, P.R. China.
authors.forename: Rui
authors.initials: R
authors.lastname: Hao
authors.affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, P.R. China.
authors.forename: Yuanchen
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Microelectronics Equipment Research and Development Center, Institute of Microelectronics of Chinese Academy of Sciences, Beijing 100029, P.R. China.
authors.forename: Chaobo
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Vascular Surgery, Clinical Division of Surgery, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, P.R. China.
authors.forename: Deyong
authors.initials: D
authors.lastname: Chen
authors.affiliation: Department of Cellular and Molecular Biology, Beijing Chest Hospital, Capital Medical University, Beijing 101149, P.R. China.
authors.forename: Xiaoting
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Vascular Surgery, Clinical Division of Surgery, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Shaoliang
authors.initials: S
authors.lastname: Luan
authors.affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, P.R. China.
authors.forename: Beiyuan
authors.initials: B
authors.lastname: Fan
authors.affiliation: Department of Vascular Surgery, Clinical Division of Surgery, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, P.R. China.
authors.forename: Junbo
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D000072283:A549 Cells; D001681:Biological Assay; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D045744:Cell Line, Tumor; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry; D015194:Diffusion Chambers, Culture; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005109:Extracellular Matrix / Q000737:chemistry; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D055616:Microtechnology / Q000295:instrumentation / Q000379:methods*; D009928:Organ Specificity; D011091:Polyesters / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry
chemical_list: D004129:Dimethylpolysiloxanes; C067625:FN1 protein, human; D005353:Fibronectins; D011091:Polyesters; D011092:Polyethylene Glycols; C033616:poly(lactide); C013830:baysilon; D003094:Collagen