A high-throughput microfluidic method for fabricating aligned collagen fibrils to study Keratocyte behavior
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A high-throughput microfluidic method for fabricating aligned collagen fibrils to study Keratocyte behavior Kevin H. Lam 1 & Pouriska B. Kivanany 2 & Kyle Grose 2 & Nihan Yonet-Tanyeri 2 & Nesreen Alsmadi 1 & Victor D. Varner 1,3 & W. Matthew Petroll 2 & David W. Schmidtke 1,3
# Springer Science+Business Media, LLC, part of Springer Nature 2019
Abstract In vivo, keratocytes are surrounded by aligned type I collagen fibrils that are organized into lamellae. A growing body of literature suggests that the unique topography of the corneal stroma is an important regulator of keratocyte behavior. In this study we describe a microfluidic method to deposit aligned fibrils of type I collagen onto glass coverslips. This high-throughput method allowed for the simultaneous coating of up to eight substrates with aligned collagen fibrils. When these substrates were integrated into a PDMS microwell culture system they provided a platform for high-resolution imaging of keratocyte behavior. Through the use of wide-field fluorescence and differential interference contrast microscopy, we observed that the density of collagen fibrils deposited was dependent upon both the perfusion shear rate of collagen and the time of perfusion. In contrast, a similar degree of fibril alignment was observed over a range of shear rates. When primary normal rabbit keratocytes (NRK) were seeded on substrates with a high density of aligned collagen fibrils and cultured in the presence of platelet derived growth factor (PDGF) the keratocytes displayed an elongated cell body that was co-aligned with the underlying collagen fibrils. In contrast, when NRK were cultured on substrates with a low density of aligned collagen fibrils, the cells showed no preferential orientation. These results suggest that this simple and inexpensive method can provide a general platform to study how simultaneous exposure to topographical and soluble cues influence cell behavior. Keywords Collagen . Fibrils . Keratocytes . Microfluidic . PDMS . Aligned
1 Introduction It is now well appreciated that biochemical, mechanical, and topographical signals emanating from the local extracellular matrix (ECM) are key modulators of cell behavior during both physiological and pathological processes. For example biochemical gradients are important in regulating cell behavior during development (Tabata 2004), inflammation (David and Kubes Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10544-019-0436-3) contains supplementary material, which is available to authorized users. * David W. Schmidtke [email protected] 1
Department of Bioengineering, University of Texas at Dallas, 800 W. Campbell Rd, Richardson, TX 75080, USA
2
Department of Ophthalmology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390-9057, USA
3
Department of Surgery, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390-9057, USA
2019), and cancer metastasis (Roussos et al. 2011), while mechanical cues are relevan
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