Single cell trapping in larger microwells capable of supporting cell spreading and proliferation |
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Authors: | Joong Yull Park Mina Morgan Aaron N Sachs Julia Samorezov Ryan Teller Ye Shen Kenneth J Pienta Shuichi Takayama |
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Affiliation: | (1) Department of Biomedical Engineering, College of Engineering, University of Michigan, 2200 Bonisteel Blvd, Ann Arbor, MI 48109, USA;(2) Departments of Internal Medicine and Urology, Michigan Center for Translational Pathology and the Comprehensive Cancer Center, University of Michigan, 1500 E. Medical Center Drive, Ann Arbor, MI 48109, USA; |
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Abstract: | Conventional cell trapping methods using microwells with small dimensions (10–20 μm) are useful for examining the instantaneous
cell response to reagents; however, such wells have insufficient space for longer duration screening tests that require observation
of cell attachment and division. Here we describe a flow method that enables single cell trapping in microwells with dimensions
of 50 μm, a size sufficient to allow attachment and division of captured cells. Among various geometries tested, triangular
microwells were found to be most efficient for single cell trapping while providing ample space for cells to grow and spread.
An important trapping mechanism is the formation of fluid streamlines inside, rather than over, the microwells. A strong flow
recirculation occurs in the triangular microwell so that it efficiently catches cells. Once a cell is captured, the cell presence
in the microwell changes the flow pattern, thereby preventing trapping of other cells. About 62% of microwells were filled
with single cells after a 20 min loading procedure. Human prostate cancer cells (PC3) were used for validation of our system. |
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