title: Automation of three-dimensional cell culture in arrayed microfluidic devices.
abstract: The increasing interest in studying the interactions between cells and the extracellular matrix (ECM) has created a need for high throughput low-cost three-dimensional (3D) culture systems. The recent development of tubeless microfluidics via passive pumping provides a high throughput microchannel culture platform compatible with existing high throughput infrastructures (e.g., automated liquid handlers). Here, we build on a previously reported high throughput two-dimensional system to create a robust automated system for 3D culture. Operational controls including temperature and sample handling have been characterized and automated. Human mammary fibroblasts (HMFs) suspended in type I collagen are loaded and cultured in microchannel arrays and used to optimize the system operational parameters. A Peltier cooler maintains the collagen as a liquid at 4 °C during cell seeding, followed by polymerization at 37 °C. Optimization of this platform is discussed (e.g., controlling collagen contraction, increasing cell viability, preventing the removal of microchannel contents), and 3D distribution of HMFs is examined by fluorescent microscopy. Finally, we validate the platform by automating a previously developed 3D breast carcinoma coculture assay. The platform allows more efficient 3D culture experiments and lays the foundation for high throughput studies of cell-ECM interactions.
authors: Materials Science Program, University of Wisconsin-Madison, Madison, WI 53705, USA.
authors: Sara I
authors: SI
authors: Montanez-Sauri
authors: Kyung Eun
authors: KE
authors: Sung
authors: John P
authors: JP
authors: Puccinelli
authors: Carolyn
authors: C
authors: Pehlke
authors: David J
authors: DJ
authors: Beebe
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