title: A microwell pattern for C17.2 cell aggregate formation with concave cylindrical surface induced cell peeling.
abstract: We have developed a polydimethylsiloxane (PDMS) pattern with arrays of microwells for the formation of multicellular aggregates by C17.2 neural stem cells. Upon interfacing with the patterns, the neural stem cells would firstly attach to the microwell sidewalls, forming cellular strips on day 1 after plating. For channel connected microwells, cellular strips on the concave semi-cylindrical sidewall surfaces continued among wells and through channels, followed by strip peeling due to prestress arising from actin filaments and assembly of suspending cellular aggregates within the microwells in the following 1-2 days. Our results also suggested that a small microwell diameter of 80 and 100 μm and a narrow channel width of 20 μm would facilitate the aggregate formation among the structural dimensions tested. Finite element method (FEM) simulation revealed that cellular strips on the semi-cylindrical sidewall surfaces peeled under significantly smaller prestresses (critical peeling prestress, CPP), than cells on flat substrates. However, the CPP by itself failed to fully account for the difference in aggregate inducing capability among the patterns addressed, suggesting cell growth behaviors might play a role. This study thus justified the current patterning method as a unique and practical approach for establishing 3D neural stem cell-based assay platforms.
authors.affiliation: Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, PR China.
authors.forename: Li-Guang
authors.initials: LG
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, PR China.
authors.forename: Dong-Huo
authors.initials: DH
authors.lastname: Zhong
authors.affiliation: Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, PR China.
authors.forename: Yiguo
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Nanobioengineering/Bioelectronics Laboratory, Department of Biomedical Engineering, Florida International University, Miami, FL 33174, USA.
authors.forename: Chen-Zhong
authors.initials: CZ
authors.lastname: Li
authors.affiliation: Cellular Bioengineering Laboratory, Faculty of Engineering, University of Georgia, Athens, GA 30602, USA.
authors.forename: William S
authors.initials: WS
authors.lastname: Kisaalita
authors.affiliation: Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, PR China. Electronic address: zzwu@cqu.edu.cn.
authors.forename: Ze-Zhi
authors.initials: ZZ
authors.lastname: Wu
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D021621:Imaging, Three-Dimensional; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D058953:Neural Stem Cells / Q000166:cytology*; D016596:Vinculin / Q000378:metabolism
chemical_list: D000199:Actins; D004129:Dimethylpolysiloxanes; D016596:Vinculin; C013830:baysilon
keywords: Cellular aggregate; Finite element analysis; Neural stem cell; Polydimethylsiloxane; Three dimensional